Methods of preventing or treating pain using anti-ngf antibodies

ABSTRACT

Antibodies and antibody fragments thereof with binding specificity to human Nerve Growth Factor (NGF) and methods of use for treating pain. Methods of treating pain or eliciting an analgesic effect comprising administering an effective amount of an anti-human NGF antibody or antibody fragment thereof, which inhibits the association of NGF with TrkA, and/or p75. These methods may optionally further comprising administering an effective amount of a second anti-human NGF antibody or fragment thereof (e.g., one which inhibits the association of NGF with p75, or one that inhibits the association of NGF with TrkA.)

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of U.S. patent application Ser. No.14/665,578 filed Mar. 23, 2015, which is a Divisional application ofU.S. patent application Ser. No. 13/834,889, filed Mar. 15, 2013, nowU.S. Pat. No. 9,067,988, which is a Continuation-in-part application ofU.S. patent application Ser. No. 13/309,153, filed Dec. 1, 2011, nowU.S. Pat. No. 8,728,473, which claims the benefit of priority to U.S.provisional patent application No. 61/418,832, filed Dec. 1, 2010, thecontents of which are incorporated herein by reference in theirentireties. In addition this application relates to U.S. patentapplication Ser. No. 13/308,665, now U.S. Pat. No. 8,911,734, entitled“METHODS OF PREVENTING OR TREATING PAIN USING ANTI-NGF ANTIBODIES THATSELECTIVELY INHIBIT THE ASSOCIATION OF NGF WITH TRKA, WITHOUT AFFECTINGTHE ASSOCIATION OF NGF WITH P75”; U.S. patent application Ser. No.13/309,831, entitled “METHODS OF PREVENTING INFLAMMATION AND TREATINGPAIN USING ANTI-NGF COMPOSITIONS”; and U.S. patent application Ser. No.13/309,295, entitled “ANTI-NGF COMPOSITIONS AND USE THEREOF”, allassigned to ALDERBIO HOLDINGS LLC, and all filed on Dec. 1, 2011, thecontents of which are all incorporated by reference in their entireties.

SEQUENCE LISTING

The sequence listing in the file named “43257o2016.txt” having a size of307,066 bytes that was created Jul. 20, 2017 is hereby incorporated byreference in its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

This invention pertains to anti-pain medicaments comprising at least oneantibody or fragment thereof (including Fab fragments) having bindingspecificity to human Nerve Growth Factor (hereinafter “NGF”), andmethods of using one or more of said antibodies and fragments thereof totreat pain in an individual. These antibodies and antibody fragments maybe used as a monotherapy to treat or prevent different types of pain ina subject in need thereof when administered alone or in association withanother active agent, e.g., a NSAID or opioid analgesic. Morespecifically the invention pertains to anti-human NGF antibodies orfragments thereof that inhibit the association of NGF with p75 and/orTrkA. In addition, and related thereto the invention pertains to novelmethods of treating pain or eliciting an analgesic effect in anindividual, comprising administering an effective amount of ananti-human NGF antibody or fragment thereof which inhibits theassociation of NGF with TrkA and/or p75.

Description of Related Art

Nerve Growth Factor (NGF) (also known as beta nerve growth factor(Beta-NGF)) is produced as a mature protein of 222 amino acids inlength, following cleavage of a 18 amino acid signal peptide. The geneencoding NGF is located on chromosome 1p13.1. A biologically active formof NGF is a secreted protein which homodimerizes and is incorporatedinto a larger complex. NGF is a member of the neurotrophins (NTs), whichare a group of structurally-related proteins further includingbrain-derived neurotrophic factor (BDNF), NT-3, and NT-4/5. (Wyman etal., Gene Therapy (1999), 6:1648-1660). NTs support the survival ofspecific types of neurons and neurotransmitter systems, being producedby cells that are targeted by innervating neurons. Id. Basal forebrain,substantia nigra, brain stem, cortex, and spinal cord are nervous systemregions having demonstrated responsiveness to NGF. Id.

All NTs bind to a low-affinity receptor identified as p75. (Sarchielliet al., Expert Rev. Neurotherapeutics (2004), 4(1):115-127). NGFselectively binds to, and displays a high affinity for, the highaffinity neurotrophin receptor TrkA. Id. It has recently beendemonstrated that NGF acts through its low-affinity receptor p75 in adevelopmentally-regulated signaling pathway necessary for myogenicdifferentiation and muscle repair in vivo. (Deponti et al., Mol. Biol.Cell (2009), 20:3620-3627).

NGF has also been demonstrated to interact with pain-signalling systemsin adult animals, and is responsible for hyperalgesia when administeredeither locally or systemically in many species. (Sarchielli et al.,Expert Rev. Neurotherapeutics (2004), 4(1):115-127). NGF has been shownto induce a pain-like response when infused into the CSF in rats, andhas been demonstrated to maintain chronic pain. Furthermore, NGF hasbeen demonstrated to contribute to the development of mechanicalallodynia occurring 8-12 hours later, and to the secondary painresponse. Id.

Pain may often be addressed through the administration of certainnarcotics or non-steroidal anti-inflammatory drugs (NSAIDs). However,the administration of these treatments may occur at the cost of certainnegative consequences. NSAIDs have the potential to cause kidneyfailure, intestinal bleeding, and liver dysfunction. Narcotics have thepotential to cause nausea, vomiting, impaired mental functioning, andaddiction. Therefore, it is desirable to identify alternative treatmentsfor pain in order to avoid certain of these negative consequences.

NGF is believed to play a role in a multitude of diseases and disorders,including but not limited to pain associated with a broad range ofdiseases and disorders, such as pain associated with cancers,neuropathic pain, and neurogenic pain. Due to the perceived involvementof NGF in a wide range of pain-related diseases and disorders, thereremains a need in the art for compositions and methods useful forpreventing or treating diseases and disorders associated with NGF, andparticularly those associated with pain. Particularly preferred anti-NGFcompositions are those having minimal or minimizing adverse reactions,such as inflammation when administered to the patient. Compositions ormethods that reduce or inhibit diseases or disorders associated withNGF, such as pain, are beneficial to the patient in need thereof.

BRIEF SUMMARY OF THE INVENTION

The present invention is directed to methods of treating pain andcompositions for use therein including antibodies and fragments thereofhaving binding specificity for NGF, in particular antibodies havingdesired epitopic specificity, high affinity or avidity and/or functionalproperties in the treatment of pain. These antibodies and antibodyfragments are used alone or in conjunction with other actives, includingbut not limited to other analgesics and biologics such as other NGFantagonists, e.g., other anti-NGF antibodies. More specifically theinvention pertains to anti-human NGF antibodies or fragments thereofthat specifically bind NGF which are used to treat and prevent pain,e.g., treatment of conditions associated with pain wherein NGF levelsare elevated. In preferred embodiments the antibodies or antibodyfragments will block or inhibit the association of NGF with TrkA and/orp75 and/or bind NGF/p75 complexes and/or bind NGF/TrkA complexes. Inaddition, and related thereto the invention is directed to novel methodsof treating pain or eliciting an analgesic effect in an individual,comprising administering an effective amount of an anti-human NGFantibody or fragment thereof that specifically bind NGF, e.g.,antibodies and antibody fragments which inhibit the association of NGFwith TrkA and/or p75 and/or which specifically bind to NGF/TrkAcomplexes and//or NGF/p75 complexes. The subject invention provides aplurality of novel high affinity anti-human NGF antibodies and fragmentsthereof that were produced against NGF. The invention in particularprovides anti-NGF antibodies and fragments thereof which inhibit theassociation of NGF with TrkA and/or p75 and/or which specifically bindNGF/TrkA complexes or NGF/−75 complexes having therapeutic potential intreating and preventing pain and pain-associated conditions identifiedinfra. These antibodies and fragments may be used as a monotherapy ormay be used in conjunctions with other actives including otheranalgesics and other biologics including other anti-NGF antibodies andantibody fragments. In addition the invention provides methods forproducing, identifying and isolating other anti-human NGF antibodies orfragments thereof. Another embodiment of this invention relates tomethods of using the antibodies described herein, comprising thesequences of the V_(H), V_(L) and CDR polypeptides described herein, andthe polynucleotides encoding them to treat or prevent pain byselectively inhibiting the association of NGF with TrkA and/or p75and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes. This invention therefore provides novel human monoclonalantibodies that are therapeutically useful for managing pain andanalgesics.

Preferably, the invention provides monoclonal antibodies that bind tonerve growth factor (NGF) that are useful in treating or preventingpain, e.g., associated with cancer, migraine, pre or post surgeryassociated pain and other pain related conditions. Preferably, themonoclonal antibodies are human monoclonal antibodies and neutralizebiological activities of NGF, including especially ameliorating theeffects of NGF-mediated pain responses, e.g., those involving the TrkAand/or p75 pathway including those which specifically bind NGF/TrkAcomplexes and/or NGF/p75 complexes. Also provided by the invention arecells that produce, and most preferably, secrete into cell culture mediathe monoclonal antibodies of the invention. In addition to their use fortreating and managing pain, the antibodies of the invention are usefulfor treating neuropathic and inflammatory pain-related responses thatinvolve the use of antibodies that bind to nerve growth factor (NGF),which inhibit the association of NGF with TrkA and/or p75 and/or whichspecifically bind NGF/TrkA complexes and/or NGF/p75 complexes.

In one aspect, the invention features a method for preventing ortreating post-surgical pain (referred to interchangeably as“post-incisional” or “post-traumatic pain”) by administering an antibodyor antibody fragment specific to NGF according to the invention thatbinds to nerve growth factor (NGF), preferably one which inhibits theassociation of NGF with TrkA, and/or p75 and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. This includes inhibiting orblocking the pain or analgesia resulting from post-surgical pain,including pain from surgery or from an incisional or traumatic wound. Inaddition the invention provides pharmaceutical compositions suitable foruse in treating or preventing said pain indications, preferably forhuman therapy, comprising an effective amount of at least one anantibody or antibody fragment specific to NGF according to the inventionthat binds to nerve growth factor (NGF), which inhibits the associationof NGF with TrkA, and/or p75. As discussed infra, these compositions maybe administrable by different routes and dosage regimens.

In another aspect, the invention provides methods for reducing incidenceof post-surgical pain, ameliorating post-surgical pain, palliatingpost-surgical pain; and/or delaying the development or progression ofpost-surgical pain in an individual, said method comprisingadministering an effective amount of an anti-NGF antibody or antibodyfragment according to the invention, preferably one that binds to nervegrowth factor (NGF), which inhibits the association of NGF with TrkA,and/or p75 and/or which specifically bind NGF/TrkA complexes and/orNGF/p75 complexes. In addition the invention provides pharmaceuticalcompositions suitable for use in treating or preventing said painindications, preferably for human therapy, comprising an effectiveamount of at least one an antibody or antibody fragment specific to NGFaccording to the invention that binds to nerve growth factor (NGF),preferably one which inhibits the association of NGF with TrkA, and/orp75 and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes. As discussed infra, these compositions may be administrableby different routes and dosage regimens.

In another aspect, the invention provides methods for increasing painthreshold in an individual comprising administering an effective amountof an anti-NGF antibody or antibody fragment according to the invention,preferably one that binds to nerve growth factor (NGF), which inhibitsthe association of NGF with TrkA, and/or p75 and/or which specificallybind NGF/TrkA complexes and/or NGF/p75 complexes. In addition theinvention provides pharmaceutical compositions suitable for use intreating or preventing said pain indications, preferably for humantherapy, comprising an effective amount of at least one an antibody orantibody fragment specific to NGF according to the invention, preferablyone that binds to nerve growth factor (NGF), which inhibits theassociation of NGF with TrkA, and/or p75 and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. As discussed infra, thesecompositions may be administrable by different routes and dosageregimens.

In another aspect, the invention provides methods for enhancing recoveryfrom surgery and/or injury-induced traumatic wound in an individualcomprising administering an effective amount of an anti-NGF antibody orantibody fragment according to the invention that binds to nerve growthfactor (NGF), preferably one which inhibits the association of NGF withTrkA, and/or p75 and/or which specifically bind NGF/TrkA complexesand/or NGF/p75 complexes. In some embodiments, resting pain issuppressed, ameliorated and/or prevented, in some embodiments,mechanically-induced pain (including pain resulting from movement) issuppressed, ameliorated and/or prevented, and in some embodiment,thermally-induced pain is suppressed, ameliorated and/or prevented. Insome embodiments, mechanically-induced pain is suppressed, amelioratedand/or prevented by administering an anti-NGF antibody or fragmentaccording to the invention that binds to nerve growth factor (NGF),which inhibits the association of NGF with TrkA, and/or p75 and/or whichspecifically bind NGF/TrkA complexes and/or NGF/p75 complexes. In someembodiments, resting pain is suppressed, ameliorated and/or prevented byadministering an anti-NGF antibody or fragment according to theinvention that binds to nerve growth factor (NGF), which inhibits theassociation of NGF with TrkA, and/or p75 and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. In some embodiment,thermally-induced pain is suppressed, ameliorated and/or prevented byadministering an anti-NGF antibody or fragment according to theinvention that binds to nerve growth factor (NGF), which inhibits theassociation of NGF with TrkA, and/or p75 and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. In some embodiments,allodynia (i.e., increased response (i.e., increased noxious sensation)to a normally non-noxious stimulus)) is suppressed, ameliorated and/orprevented, and/or hyperalgesia (i.e., increased response to a normallynoxious or unpleasant stimulus) is suppressed, ameliorated and/orprevented. In still further embodiments, allodynia and/or hyperalgesiais thermal or mechanical (tactile) in nature, or resting pain. In someembodiments, the pain is chronic pain. In other embodiments, the pain isassociated with site of incision, wound or trauma, and/or proximal, ator near the site of incision, wound, and/or trauma. In addition theinvention provides pharmaceutical compositions suitable for use intreating or preventing said pain indications, preferably for humantherapy, comprising an effective amount of at least one an antibody orantibody fragment specific to NGF according to the invention that bindsto nerve growth factor (NGF), which inhibits the association of NGF withTrkA, and/or p75. As discussed infra, these compositions may beadministrable by different routes and dosage regimens.

Another aspect of the invention consists in using the capacity of anantibody or antibody fragment according to the invention that binds tonerve growth factor (NGF), which inhibits the association of NGF withTrkA, and/or p75 and/or which specifically bind NGF/TrkA complexesand/or NGF/p75 complexes to bring relief to the patient suffering fromchronic visceral pain. The subject antibodies or antibody fragments arecapable of inhibiting or blocking the visceral hypersensitivity presentin the pathophysiology of visceral functional disorders, in the case ofchronic pain. Herein, the expression chronic visceral functionaldisorders is understood to include by way of example disorders of thesensitivity of the viscera having a nervous origin, also known by thename visceralgia. The viscera include the digestive, respiratory andurogenital organs and the endocrine systems, as well as the spleen, theheart and the large vessels. From the medical point of view, a chronicvisceralgia is characterized by a threshold of sensitivity to pain whichis lowered compared with the normal threshold, in response to externalmechanical stimuli. Chronic visceral pain is in addition characterizedby the absence of an inflammatory situation concomitant with thefunctional disorders. More specifically, chronic visceral pain includesthe following chronic disorders: chronic dyspepsia, a functionaldigestion disorder occurring in the absence of a detectable organiclesion and which may be symptomatic of other diseases or otherdisorders; chronic dysmenorrhea, characterized by pain associated withmenstruation; chronic pancreatitis, which is characterized by rapid lossof weight, asthenia, pain at the pancreatic point, a jaundice withdistension of the gall bladder and digestive disorders due to pancreaticinsufficiency, including hereditary chronic pancreatitis, a dominantautosomally transmitted disease which manifests itself from childhood byabdominal and recidivous painful attacks and which is characterized inadults by signs of insufficiency as well as by calcifications of thepancreas; chronic gastroesophageal reflux, which is characterized by areturn into the esophagus of the acidic gastric content and whichcauses, generally after a meal, ascending retrosternal burns, sometimesaccompanied by acidic regurgitations; IBS (irritable bowel syndrome),which is a non-inflammatory chronic disease characterized by abdominalpain and diarrhea and/or constipation, with no detectable biochemicaland histological modification. In addition the invention providespharmaceutical compositions suitable for use in treating or preventingsaid pain indications, preferably for human therapy, comprising aneffective amount of at least one an antibody or antibody fragmentspecific to NGF according to the invention that binds to nerve growthfactor (NGF), which inhibits the association of NGF with TrkA, and/orp75 and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes. As discussed infra, these compositions may be administrableby different routes and dosage regimens.

These antibodies and antibody fragments may be used to treat differentconditions associated with pain such as are identified herein includingby way of example IBS and chronic visceral pain, in particulargastrointestinal pain. The subject antibodies or fragments according tothe invention may be used for the manufacture of a medicament intendedfor the prevention or treatment of chronic visceral pain or any disorderor condition involving NGF-associated pain.

In another embodiment, the invention provides methods for enhancingopioid analgesic pain treatment comprising administering an effectiveamount of an opioid analgesic in conjunction with an effective amount ofan anti-NGF antibody or antibody fragment according to the invention,preferably one that binds to nerve growth factor (NGF), which inhibitsthe association of NGF with TrkA, and/or p75 and/or which specificallybind NGF/TrkA complexes and/or NGF/p75 complexes. Administration inconjunction, as used herein, comprises simultaneous administrationand/or administration at different times. Administration in conjunctionalso encompasses administration as a co-formulation (i.e., the NGFantibody or fragment according to the invention and opioid analgesic arepresent (combined) in the same composition) and/or administration asseparate compositions. As used herein, “administration in conjunction”is meant to encompass any circumstance wherein an NGF antibody orfragment according to the invention, preferably one that binds to nervegrowth factor (NGF), which inhibits the association of NGF with TrkA,and/or p75 and/or which specifically bind NGF/TrkA complexes and/orNGF/p75 complexes and another active, e.g. another analgesic agent areadministered in an effective amount to an individual. In addition theinvention provides pharmaceutical compositions suitable for use intreating or preventing said pain indications, preferably for humantherapy, comprising an effective amount of at least one an antibody orantibody fragment specific to NGF according to the invention that bindsto nerve growth factor (NGF), and which preferably inhibits theassociation of NGF with TrkA, and/or p75 and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. As discussed infra, thesecompositions may be administrable by different routes and dosageregimens.

As further discussed herein, it is understood that the anti-NGF antibodyor fragment and other active, e.g., opioid analgesic can be administeredat different dosing frequencies and/or intervals. For example, ananti-NGF antibody or fragment can be administered weekly, while anopioid analgesic can be administered more frequently. It is understoodthat the NGF antibody or fragment and the opioid analgesic can beadministered using the same route of administration or different routesof administration, and that different dosing regimens may change overthe course of administration(s). Administration may be before the onsetof pain.

In another aspect, the invention provides methods for reducing incidenceof pain, ameliorating pain, palliating pain; and/or delaying thedevelopment or progression of pain in an individual, said methodscomprising administering an effective amount of an opioid analgesic inconjunction with an effective amount of an NGF antibody or fragmentaccording to the invention that binds to nerve growth factor (NGF),preferably one which inhibits the association of NGF with TrkA, and/orp75 and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes. The methods of the invention are suitable for treating orpreventing any pain of any etiology, including pain where the use of anopioid analgesic is generally prescribed, for example, pancreatitis,kidney stone, headache, dysmenorrhea, musculoskeletal pain (e.g., lowerback pain), sprain, visceral pain, ovarian cysts, prostatitis, cystitis,chemical or thermal burns, or cancer (such as prostate cancermetastasized to bone, breast cancer that has metastasized to bone, lungcancer that has metastasized to bone, pancreatic cancer). In additionthe invention provides pharmaceutical compositions suitable for use intreating or preventing said pain indications, preferably for humantherapy, comprising an effective amount of at least one an antibody orantibody fragment specific to NGF according to the invention that bindsto nerve growth factor (NGF), preferably one which inhibits theassociation of NGF with TrkA, and/or p75. As discussed infra, thesecompositions may be administrable by different routes and dosageregimens.

In another aspect, the present invention features a method for treating(or, in other embodiments, preventing) pain comprising administering anamount of an anti-NGF antibody or fragment according to the invention inassociation with another active, e.g., another analgesic agent in orderto provide effective pain relief. In addition the invention providespharmaceutical compositions suitable for use in treating or preventingsaid pain indications, preferably for human therapy, comprising aneffective amount of at least one an antibody or antibody fragmentspecific to NGF according to the invention that binds to nerve growthfactor (NGF), which inhibits the association of NGF with TrkA, and/orp75 and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes. As discussed infra, these compositions may be administrableby different routes and dosage regimens.

In another specific aspect, the invention provides methods for enhancingNSAID pain treatment comprising administering an effective amount of anNSAID in conjunction with an effective amount of an anti-anti-NGFantibody or antibody fragment according to the invention that binds tonerve growth factor (NGF), preferably one which inhibits the associationof NGF with TrkA, and/or p75 and/or which specifically bind NGF/TrkAcomplexes and/or NGF/p75 complexes. Administration in conjunction, asused herein, comprises simultaneous administration and/or administrationat different times. Administration in conjunction also encompassesadministration as a co-formulation (i.e., the NGF antibody and NSAID arepresent (combined) in the same composition) and/or administration asseparate compositions. As used herein, “administration in conjunction”is meant to encompass any circumstance wherein an NSAID and anti-NGFantibody are administered in an effective amount to an individual. Asfurther discussed herein, it is understood that the NGF antibody orfragment and NSAID can be administered at different dosing frequenciesand/or intervals. For example, an anti-NGF antibody can be administeredweekly, while an NSAID can be administered more frequently. It isunderstood that the NGF antibody and the NSAID can be administered usingthe same route of administration or different routes of administration,and that different dosing regimens may change over the course ofadministration(s). Administration may be before the onset of pain. Inaddition the invention provides pharmaceutical compositions suitable foruse in treating or preventing said pain indications, preferably forhuman therapy, comprising an effective amount of at least one anantibody or antibody fragment specific to NGF according to the inventionthat binds to nerve growth factor (NGF), which inhibits the associationof NGF with TrkA, and/or p75 and/or which specifically bind NGF/TrkAcomplexes and/or NGF/p75 complexes. As discussed infra, thesecompositions may be administrable by different routes and dosageregimens.

In another aspect, the invention provides methods for reducing incidenceof pain, ameliorating pain, palliating pain, and/or delaying thedevelopment or progression of pain in an individual, said methodscomprising administering an effective amount of an NGF antibody orantibody fragment according to the invention that binds to nerve growthfactor (NGF), preferably one which inhibits the association of NGF withTrkA, and/or p75 and/or which specifically bind NGF/TrkA complexesand/or NGF/p75 complexes in conjunction with an effective amount of anNSAID. The methods of the invention are suitable for treating orpreventing any pain of any etiology, including pain where the use of anNSAID is generally prescribed. In some embodiments, the pain ispost-surgical pain. In some embodiments, the pain is pain associatedwith burn. In other embodiments, the pain is pain associated withrheumatoid arthritis. In other embodiments, the pain is pain associatedwith osteoarthritis. In addition the invention provides pharmaceuticalcompositions suitable for use in treating or preventing said painindications, preferably for human therapy, comprising an effectiveamount of at least one an antibody or antibody fragment specific to NGFaccording to the invention that binds to nerve growth factor (NGF),preferably one which inhibits the association of NGF with TrkA, and/orp75 and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes. As discussed infra, these compositions may be administrableby different routes and dosage regimens.

In another aspect, the invention features a method for preventing ortreating bone cancer pain including cancer pain associated with bonemetastasis (also termed “bone metastasis pain”) by administering ananti-NGF antibody or antibody fragment according to the invention thatbinds to nerve growth factor (NGF), preferably one which inhibits theassociation of NGF with TrkA, and/or p75 and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. In some embodiments, theNGF antibody or fragment is co-administered with an opioid analgesic. Insome embodiments, the NGF antibody or antibody fragment according to theinvention is co-administered with an NSAID. In some embodiments, the NGFantagonist is co-administered with an opioid analgesic and an NSAID. Insome embodiments, the NGF antagonist is not co-administered with anopioid analgesic. In some embodiments, the NGF antagonist is notco-administered with an NSAID. In addition the invention providespharmaceutical compositions suitable for use in treating or preventingsaid pain indications, preferably for human therapy, comprising aneffective amount of at least one an antibody or antibody fragmentspecific to NGF according to the invention that binds to nerve growthfactor (NGF), which inhibits the association of NGF with TrkA, and/orp75. As discussed infra, these compositions may be administrable bydifferent routes and dosage regimens.

In another aspect, the invention provides methods for reducing incidenceof bone cancer pain including cancer pain associated with bonemetastasis, ameliorating bone cancer pain including cancer painassociated with bone metastasis, palliating bone cancer pain includingcancer pain associated with bone metastasis; and/or delaying thedevelopment or progression of bone cancer pain including cancer painassociated with bone metastasis in an individual, said methodscomprising administering an effective amount of an NGF antibody orfragment according to the invention that binds to nerve growth factor(NGF), preferably one which inhibits the association of NGF with TrkA,and/or p75 and/or which specifically bind NGF/TrkA complexes and/orNGF/p75 complexes. In some embodiments, the NGF antibody or fragment isco-administered with an opioid analgesic. In some embodiments, the NGFantibody or fragment is co-administered with an NSAID. In someembodiments, the NGF antibody or fragment is co-administered with anopioid analgesic and an NSAID. In some embodiments, the NGF antibody orfragment is not co-administered with an opioid analgesic. In someembodiments, the NGF antibody or fragment is not co-administered with anNSAID. In addition the invention provides pharmaceutical compositionssuitable for use in treating or preventing said pain indications,preferably for human therapy, comprising an effective amount of at leastone an antibody or antibody fragment specific to NGF according to theinvention that binds to nerve growth factor (NGF), preferably one whichinhibits the association of NGF with TrkA, and/or p75 and/or whichspecifically bind NGF/TrkA complexes and/or NGF/p75 complexes. Asdiscussed infra, these compositions may be administrable by differentroutes and dosage regimens.

In some embodiments, the bone cancer pain is from cancer originated inbone. In some embodiments, the bone cancer pain is from osteosarcoma. Insome embodiments, the bone cancer pain is from cancer metastasized tobone. In some embodiments, the bone metastasis is prostate cancermetastasized to bone. In some embodiments, the bone metastasis is breastcancer metastasized to bone. In some embodiments, the bone metastasis islung cancer metastasized to bone. In some embodiments, the bonemetastasis is sarcoma metastasized to bone. In some embodiments, thebone metastasis is kidney cancer metastasized to bone. In someembodiments, the bone metastasis is multiple myeloma metastasized tobone. In some embodiments, the cancer pain treated is mild to moderate.In some embodiments, the cancer pain treated is moderate to severe. Insome embodiments, the cancer pain treated is severe.

In preferred embodiments of the invention methods of treating pain areprovided using chimeric or humanized antibodies and fragments thereofcapable of binding to NGF and preferably selectively inhibiting thebiological activities mediated by the binding of NGF to the TrkAreceptor, while not inhibiting the biological activities mediated by thebinding of NGF to the p75 receptor and/or which specifically bindNGF/TrkA complexes and/or NGF/p75 complexes. In another preferredembodiment of the invention, full length antibodies and Fab fragmentsthereof are provided which bind to nerve growth factor (NGF), whichinhibit the association of NGF with TrkA, and/or p75 and/or whichspecifically bind NGF/TrkA complexes and/or NGF/p75 complexes, and thatare capable of significantly reducing pain in vivo in murine models, asmeasured by Gait analysis (as described in the examples herein) or byinhibiting PC12 neurite outgrowth as described infra. As noted theseantibodies and antibody fragments may be administered alone or inassociation with other actives such as NSAIDs or opioid analgesics. Inaddition the invention provides pharmaceutical compositions suitable foruse in treating or preventing said pain indications, preferably forhuman therapy, comprising an effective amount of at least one anantibody or antibody fragment specific to NGF according to the inventionthat binds to nerve growth factor (NGF), which inhibits the associationof NGF with TrkA, and/or p75 and/or which specifically bind NGF/TrkAcomplexes and/or NGF/p75 complexes. As discussed infra, thesecompositions may be administrable by different routes and dosageregimens.

In another embodiment of the invention, chimeric or humanized antibodiesand fragments thereof (including Fab fragments) are provided which arecapable of binding to NGF that inhibit the association of NGF with TrkA,and/or p75 and which further inhibit TF1 cell proliferation. In anotherembodiment of the invention, chimeric or humanized antibodies andfragments thereof (including Fab fragments) capable of binding to NGF,which preferably inhibits the association of NGF with TrkA, and/or p75and/or which specifically bind NGF/TrkA complexes and/or NGF/p75complexes and further which inhibit PC-12 neurite outgrowth. In additionthe invention provides pharmaceutical compositions suitable for use intreating or preventing said pain indications, preferably for humantherapy, comprising an effective amount of said chimeric or humanizedantibodies and fragments thereof (including Fab fragments) are providedwhich are capable of binding to NGF that inhibit the association of NGFwith TrkA, and/or p75 and/or which specifically bind NGF/TrkA complexesand/or NGF/p75 complexes and which preferably further inhibit TF1 cellproliferation. As discussed infra, these compositions may beadministrable by different routes and dosage regimens.

In another embodiment of the invention these antibodies and humanizedversions may be derived from rabbit immune cells (B lymphocytes) and maybe selected based on their homology (sequence identity) to human germline sequences. These antibodies may require minimal or no sequencemodifications, thereby facilitating retention of functional propertiesafter humanization. A further embodiment of the invention is directed tofragments from anti-NGF antibodies encompassing V_(H), V_(L) and CDRpolypeptides, e.g., derived from rabbit immune cells and thepolynucleotides encoding the same, as well as methods of using theseantibody fragments and the polynucleotides encoding them in the creationof novel antibodies and polypeptide compositions capable of binding toNGF and/or NGF/p75 and NGF/TrkA complexes.

The invention also contemplates conjugates of anti-NGF antibodies andbinding fragments thereof that bind to nerve growth factor, whichinhibit the association of NGF with TrkA, and/or p75 and/or whichspecifically bind NGF/TrkA complexes and/or NGF/p75 complexes, which areconjugated to one or more functional or detectable moieties. Theinvention also contemplates methods of making said chimeric or humanizedanti-NGF, anti-NGF/p75 complex or anti-NGF/TrkA complex antibodies andbinding fragments thereof. In one embodiment, binding fragments include,but are not limited to, Fab, Fab′, F(ab′)₂, Fv, scFv fragments, SMIPs(small molecule immunopharmaceuticals), camelbodies, nanobodies, MetMablike monovalent and divalent agents monospecific or bispecific innature, and IgNAR. Examples of monovalent agents include Fab, Fab′, Fv,scFv fragments, SMIPs (small molecule immunopharmaceuticals),camelbodies, nanobodies, IgNAR, a monovalent antibody molecule analogousto MetMab, or one or more combinations thereof.

The present invention includes in particular monovalent antibodymolecules that bind NGF, which are analogous to MetMab molecules. MetMabis a monovalent antibody specific to Met. (Met is a protein encoded bythe nucleotide sequence set forth in Park et al., Proc. Natl. Acad. Sci.84, 7479-(1987), or fragments thereof, as well as related polypeptides,which include, but are not limited to, allelic variants, splicevariants, derivative variants, substitution variants, deletion variants,and/or insertion variants, fusion polypeptides, and interspecieshomologs). The MetMab antibody, is a monovalent antibody known bydifferent names including OA-5d5 (Genentech) and is also called OneArmed 5d5, 5d5, MetMab, PRO143966, among others). Antibody OA-5d5,including its structure and properties, and methods for making and usingit, are described in U.S. Publication No. 2007/0092520. In oneembodiment, an anti-NGF antibody according to the invention may comprisea single Fab region linked to an Fc region. In such embodiment, anantibody of the invention may comprise light and heavy chain variabledomains as described herein. In such an embodiment, the antibody ismonovalent and may comprise an intact Fc region. In another suchembodiment, the Fc region may comprise at least one protuberance (knob)and at least one cavity (hole), wherein the presence of the protuberanceand cavity enhances formation of a complex between an Fc polypeptidecomprising the protuberance and an Fc polypeptide comprising the cavity,for example as described in WO 2005/063816. In one embodiment, the Fcregion of an antibody of the invention may comprise a first and a secondFc polypeptide, wherein the first and second polypeptide each comprisesone or more mutations with respect to wild type human Fc. In oneembodiment, a cavity mutation is T366S, L368A and/or Y407V. In anotherembodiment, a protuberance mutation is T366W. In a specific embodiment,a monovalent antibody according to the subject invention may comprise aone-armed antibody synthesized as described in WO2005/063816. In suchembodiment, the one-armed antibody may comprise Fc mutationsconstituting “knobs” and “holes” as described in WO2005/063816. Forexample, a hole mutation can be one or more of T366A, L368A and/or Y407Vin an Fc polypeptide, and a cavity mutation can be T366W.

The invention is also directed to an anti-human NGF monovalent agentthat binds with the same NGF epitope and/or competes with an anti-NGFantibody for binding to NGF as an antibody or antibody fragmentdisclosed herein, including but not limited to an anti-NGF antibodyselected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11,Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20 or Ab21.

In addition the invention provides pharmaceutical compositions suitablefor use in treating or preventing said pain indications, preferably forhuman therapy, comprising an effective amount of said conjugates ofanti-NGF antibodies and binding fragments thereof that bind to nervegrowth factor, which inhibit the association of NGF with TrkA, and/orp75, which are conjugated to one or more functional or detectablemoieties.

Embodiments of the invention pertain to the use of anti-NGF antibodiesand binding fragments thereof which inhibit the association of NGF withTrkA, and/or p75 and/or which specifically bind NGF/TrkA complexesand/or NGF/p75 complexes, for the diagnosis, assessment and treatment ofdiseases and disorders associated with NGF or aberrant expressionthereof, especially pain associated disorders. The invention alsocontemplates the use of fragments of anti-NGF antibodies which inhibitthe association of NGF with TrkA, and/or p75 and/or which specificallybind NGF/TrkA complexes and/or NGF/p75 complexes, for the diagnosis,assessment and treatment of diseases and disorders associated with NGFor aberrant expression thereof. Other embodiments of the inventionrelate to the production of anti-NGF antibodies or fragments thereofwhich inhibit the association of NGF with TrkA, and/or p75, inrecombinant host cells, for example mammalian cells such as CHO, NSO orHEK 293 cells, or yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. In addition the invention providespharmaceutically acceptable compositions for use in said methods.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIGS. 1A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab1.

FIGS. 2A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab2.

FIGS. 3A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab3.

FIGS. 4A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab4.

FIGS. 5A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab5.

FIGS. 6A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab6.

FIGS. 7A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab7.

FIGS. 8A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab8.

FIGS. 9A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab9.

FIGS. 10A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab10.

FIGS. 11A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab11.

FIGS. 12A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab12.

FIGS. 13A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab13.

FIGS. 14A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab14.

FIGS. 15A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab15.

FIGS. 16A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab16.

FIGS. 17A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab17.

FIGS. 18A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab18.

FIGS. 19A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab19.

FIGS. 20A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab20.

FIGS. 21A-B provide polynucleotide and polypeptide sequencescorresponding to the full-length Antibody Ab21, produced by expressionin Pichia pastoris.

FIG. 22 provides the heavy and light chain polypeptide sequences ofFab1.

FIG. 23 provides the heavy and light chain polypeptide sequences ofFab2.

FIG. 24 provides the NGF ELISA binding data obtained following theprotocol described infra for antibodies Ab1 and Ab2.

FIG. 25 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab3.

FIG. 26 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab4.

FIG. 27 provides the NGF ELISA binding data obtained following theprotocol described infra for antibodies Ab5 and Ab6.

FIG. 28 provides the NGF ELISA binding data obtained following theprotocol described infra for Fab1.

FIG. 29 provides the NGF ELISA binding data obtained following theprotocol described infra for Fab2.

FIG. 30 provides the NGF ELISA binding data obtained following theprotocol described infra for antibodies Ab7 and Ab8.

FIG. 31 provides the NGF ELISA binding data obtained following theprotocol described infra for antibodies Ab9 and Ab10.

FIG. 32 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab11.

FIG. 33 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab12.

FIG. 34 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab13.

FIG. 35 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab14.

FIG. 36 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab15.

FIG. 37 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab16.

FIG. 38 provides the NGF ELISA binding data obtained following theprotocol described infra for antibodies Ab17 and Ab18.

FIG. 39 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab19.

FIG. 40 provides the NGF ELISA binding data obtained following theprotocol described infra for antibody Ab20.

FIG. 41 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab1 and Ab2.

FIG. 42 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab3 and Ab4.

FIG. 43 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab5 and Ab6.

FIG. 44 provides the TF1 cell proliferation data obtained followingexample 1 for the Fab1 and Fab2 antibody fragments.

FIG. 45 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab7 and Ab8.

FIG. 46 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab9 and Ab10.

FIG. 47 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab11 and Ab12.

FIG. 48 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab13 and Ab14.

FIG. 49 provides the TF1 cell proliferation data obtained followingexample 1 for antibody Ab15.

FIG. 50 provides the TF1 cell proliferation data obtained followingexample 1 for antibody Ab16.

FIG. 51 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab17 and Ab18.

FIG. 52 provides the TF1 cell proliferation data obtained followingexample 1 for antibodies Ab19 and Ab20.

FIG. 53 provides the inhibition of NGF-p75 interaction data obtainedfollowing example 5 for antibodies Ab3 and Ab4. Antibodies Ab3 and Ab4do not demonstrate the ability to inhibit the interaction of NGF andp75.

FIG. 54 provides the inhibition of NGF-p75 interaction data obtainedfollowing example 5 for antibodies Ab15 and Ab16. Antibodies Ab15 andAb16 do not demonstrate the ability to inhibit the interaction of NGFand p75.

FIG. 55 provides the inhibition of NGF-p75 interaction data obtainedfollowing example 5 for antibody Ab5. Antibody Ab5 demonstrates theability to inhibit the interaction of NGF and p75.

FIG. 56 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab1 obtained followingexample 6.

FIG. 57 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab2 obtained followingexample 6.

FIG. 58 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab3 obtained followingexample 6. The results further show that inhibition at the same antibodyconcentrations is less than that seen with antibodies which exhibitdifferent NGF binding selectivity (i.e., compared to those which inhibitboth NGF/TrkA and NGF/p75 interactions).

FIG. 59 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab5 obtained followingexample 6.

FIG. 60 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab6 obtained followingexample 6.

FIG. 61 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab7 obtained followingexample 6.

FIG. 62 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab8 obtained followingexample 6.

FIG. 63 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab9 obtained followingexample 6.

FIG. 64 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab10 obtainedfollowing example 6.

FIG. 65 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab11 obtainedfollowing example 6.

FIG. 66 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab13 obtainedfollowing example 6.

FIG. 67 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab17 obtainedfollowing example 6.

FIG. 68 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab18 obtainedfollowing example 6.

FIG. 69 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab19 obtainedfollowing example 6.

FIG. 70 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibodies Ab2,Ab6, and Ab8, when compared with results obtained with the controlsfollowing example 7.

FIG. 71 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab6 andFab1, when compared with results obtained with the controls followingexample 7.

FIG. 72 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab3, whencompared with results obtained with the controls following example 7.

FIG. 73 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab6 andantibody Ab21, when compared with results obtained with the controlsfollowing example 7.

FIG. 74 demonstrates an increase in inflammation followingadministration of each of antibodies Ab2, Ab6, and Ab8, when comparedwith inflammation results for the controls following example 8.

FIG. 75 demonstrates no significant increase in inflammation followingadministration of the Fab1 antibody fragment, when compared withinflammation results for the control. In contrast, administration ofantibody Ab6 resulted in increased inflammation, when compared withinflammation results for the controls following example 8.

FIG. 76 demonstrates an increase in inflammation followingadministration of antibody Ab3, when compared with inflammation resultsfor the controls following example 8.

FIG. 77 also demonstrates an increase in inflammation followingadministration of antibody Ab6 and antibody Ab21, when compared withinflammation results for the controls following example 8.

FIG. 78 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab16 obtainedfollowing example 6. The results further show that inhibition with Ab16at the same antibody concentrations is less than that seen with otherantibodies which exhibit different NGF binding selectivity (i.e.,compared to those which inhibit both NGF/TrkA and NGF/p75 interactions).

FIG. 79 demonstrates the inhibition of PC-12 neurite outgrowth in thepresence of increasing concentrations of antibody Ab15 obtainedfollowing example 6. The results further show that inhibition with Ab15at the same antibody concentrations is less than that seen withantibodies which exhibit different NGF binding selectivity (i.e.,compared to those which inhibit both NGF/TrkA and NGF/p75 interactions).

FIG. 80 demonstrates no significant change in overall wellness, asdetermined by body weight, following administration of antibody Ab3 orAb15, when compared with the change in body weight for the noreactivation control. In contrast, administration of negative controlantibody resulted in a reduction in body weight, when compared with thechange in body weight for the no reactivation control.

FIG. 81 demonstrates a statistically significant reduction in pain at 72hours post-reactivation as assessed by Gait analysis followingadministration of antibody Ab3 or antibody Ab15, when compared withresults obtained with the controls following example 9.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS Definitions

It is to be understood that this invention is not limited to theparticular methodology, protocols, cell lines, animal species or genera,and reagents described, as such may vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to limit the scope ofthe present invention which will be limited only by the appended claims.As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the protein” includes reference to one or more proteinsand equivalents thereof known to those skilled in the art, and so forth.All technical and scientific terms used herein have the same meaning ascommonly understood to one of ordinary skill in the art to which thisinvention belongs unless clearly indicated otherwise.

Nerve Growth Factor (NGF): As used herein, NGF (also referred to asBeta-NGF; HSAN5; and NGFB) encompasses not only the following matureamino acid sequence available from R&D Systems (Minneapolis, Minn.) asHomo sapiens Beta-Nerve Growth Factor (β-NGF):

SSSHPIFHRGEFSVCDSVSVWVGDKTTATDIKGKEVMVLGEVNINNSVFKQYFFETKCRDPNPVDSGCRGIDSKHWNSYCTTTHTFVKALTMDGKQAAWRFIRI DTACVCVLSRKAVRRA(SEQ ID NO: 411), but also any pro-, mature, soluble, and/ormembrane-bound forms of this NGF amino acid sequence, as well as mutants(muteins), splice variants, isoforms, ortholog, homologues and variantsof this sequence.

“Pain” as used herein refers to pain of any etiology, including acuteand chronic pain, and any pain with an inflammatory component. As usedherein, “pain” includes nociception and the sensation of pain, and paincan be assessed objectively and subjectively, using pain scores andother methods well-known in the art. The pain can be primary orsecondary pain, as is well-known in the art. Deagle's Law of theMultilevel Pain Gate states, “Changes at one level of the multilevelgate cause pathway and molecular neurotransmitter and inflammatorymediator changes at all other levels.” Pain can arise at any level ofthe gating process, with different molecules responsible, and differentgenetic, pharmaceutical, immunopharmaceutical and technical challengesto block the gate mediators. Pain of all types arises through themultilevel gate, with changes in molecular upregulation at all othercontinuous levels. Thus levels of molecules that open the gate in theskin are increased in fibromyalgia and skin levels of cytokines andexcitatory neurotransmitters are increased in the brain and skin inspinal cord and peripheral nerve injuries. Pain is a neurologic signaldisease and may or may not be associated with tissue inflammation. Lackof structural disease or tissue inflammation in conditions such asfibromyalgia or other non-inflammatory pain disorders such asnon-inflammatory neuropathy, are primarily due to molecular increasedpain signal transmission. Irrespective of the type of pain, whether itis acute pain as in a sprain, sports injury or jellyfish sting, orwhether it is chronic pain as in arthritis, migraine pain, back or neckpain from herniated disks, RSD/CRPS pain, migraine, fibromyalgia,interstitial cystitis, neuropathic pain, post-stroke pain, etc., theunderlying basis is the opening of the pain gate to allow thetransmission of the pain signal from the primary or higher gates to thecortex at gate levels six and seven, where higher receptive, motor andbehavioral responses occur. This model differs substantially from themodel that inflammation is the sole cause of pain and that othermolecular modulators of pain signal transmission or inhibition are notprimary in the cause of chronic or acute pain disorders. Infibromyalgia, non-inflammatory neuromodulatory molecules are increasedin the skin as well as in other disorders at the pain gates inflammatorymolecules and profacilatory pain signal modulators are increased.Receptor density on the nerve fibers increases due to increased nervetraffic transmitting painful stimuli. Blockade of facilitativeneurotransmitters and inflammatory gate controlling molecules narrowsthe twelve-lane superhighway facilitated for pain in the chronic painstate down to a two lane normal pathway, and reduces the receptordensity for neurotransmitters as well.

This process involves pain gate facilitatory molecules. In other tissuesand circumstances, these may be proinflammatory, or neutral, oranti-inflammatory. Irrespective of the characteristic of the pain,whether it is sharp, dull, aching, burning, stabbing, numbing ortingling, all pain arises with the allowance of increased signal of painat one of the seven pain gate levels at the specific site of thestructures that comprise the gates. In the skin this structure is theLangerhans dermal structures, next the dorsal root ganglion, the dorsalhorn substantia gelatinosa, ascending spinothalamic tracts, subthalamicnuclei, sensory receptive cortex, and anterior motor and behavioralresponse cortex. Pain gate facilatatory molecules such as in thecondition of fibromyalgia can be identified in the skin with increasedfrequency in this condition. Lack of pain gate inhibitory molecules mayalso be identified at all seven levels of the gate, with upregulation ofgenes, proteins, enzymes and glial cell transformation to open the gatein pain and down-regulation of genes, proteins, enzymes and blockade ofglial cell transformation molecules in the pain state. Thus pain can beviewed as a signal disease and a neurological informational disorder,with excessive transmission to higher levels from one gate to anothergate of pain signal increasingly processed, with lack of signalinhibition until there is a cortical response. According to a study onneuropharmaceutical multilevel neural pain gate andtransduction-transmission blockades, the neural process of pain requiresmultilevel hierarchical integrated gating and requires the pain gate tobe opened by inflammatory molecules. These include interleukin 1 Beta,IL1B; necrosis factor alpha, TNF-Alpha; interleukin 6, IL-6; interleukin8, IL-8; interleukin 2, IL-2; and vanilloid receptor 1, VR1; andgranulocyte inhibitor factor, GIF; iNO, glial cell activation, and freeradical molecules such as hydroxyl OH, and nitric oxide or nitroperoxylradicals NO, and NOOH. Pain associated therewith potentially may beblocked with pharmaceuticals such as biologics including peptides andantibodies, small molecules and the like that inhibit theafore-identified cytokines, receptors or radicals.

Pain may be present in inflammatory or non-inflammatory conditions andtherefore the presence of molecules that are found to cause or propagatepain if these molecules are not found at the pain gate. Non-inflammatoryconditions such as fibromyalgia, myofascial pain, and non-inflammatorymetabolic neuritic pain syndromes are excellent examples.

For example, “pain associated with chronic prostatitis and/or chronicpelvic pain syndrome” as used herein refers to lower abdominal (pelvic)pain; lower stomach pain; bladder pain; suprapubic pain; pain in thepenis, testicles, scrotum and perineum; urethral pain; dyspareunia;pain, pressure or discomfort that may increase as the bladder fills;dysuria; and ejaculatory pain.

Although analgesia in the strictest sense is an absence of pain, as usedherein, “analgesia” refers to reduction in pain perceived by anindividual.

“Analgesia agent”, “analgesic agent” or “analgesic” refers to anybiomolecule, drug or active agent that alleviates or prevents pain. Thisincludes in particular the subject NGF antibodies and fragments as wellas other actives such as opioids and NSAIDs.

“Acute pain” refers to sudden pain from a specific cause (injury,infection, inflammation, etc.) that has lasted for a limited period oftime (as opposed to chronic pain).

“Chronic pain” refers to a persistent state of pain. Chronic pain isoften associated with long-term incurable or intractable medicalconditions or diseases.

“Procedural pain” refers to pain arising from a medical, dental surgicalor other procedure wherein the procedure may be planned or associatedwith acute trauma.

“Headache disorder” includes migraine, tension headache, clusterheadache, trigeminal neuralgia, secondary headaches, andmiscellaneous-type headaches.

“Migraine” includes migraine headache, migraine without aura, migrainewith aura, and migraine with aura but without headache.

“Systemic side effects” sometimes associated with pain include, but arenot limited to, cardiovascular including peripheral vasodilatation andinhibition of baroreceptors; dermatologic including pruritis (itching),flushing and red eyes; gastrointestinal including nausea and vomiting,decreased gastric motility, decreased biliary, pancreatic and intestinalsecretions and delays in food digestion, diminished peristaltic waves inlarge intestine contributing to constipation, epigastric distress orbiliary colic in biliary tract; respiratory including depressedrespiratory effort; and urinary including urinary urgency and difficultywith urination; and peripheral limb heaviness.

“Central nervous system side effects” or “CNS side effects” associatedwith pain sometimes include, but are not limited to, narcosis, euphoria,drowsiness, apathy, psychotic ideation, mental confusion, alteration inmood, reduction in body temperature, feelings of relaxation, dysphoria(an emotional state characterized by anxiety, depression, or unease),and nausea and vomiting (caused by direct stimulation of chemoreceptorsin the medulla).

As used herein, “treatment” is an approach for obtaining beneficial ordesired clinical results. For purposes of this invention, beneficial ordesired clinical results include, but are not limited to, one or more ofthe following: improvement or alleviation of any aspect of pain,including acute, chronic, inflammatory, neuropathic, or post-surgicalpain. For purposes of this invention, beneficial or desired clinicalresults include, but are not limited to, one or more of the following:including lessening severity, alleviation of one or more symptomsassociated with pain including any aspect of pain (such as shorteningduration of pain, and/or reduction of pain sensitivity or sensation).

“Reducing incidence” of pain means any of reducing severity (which caninclude reducing need for and/or amount of (e.g., exposure to) otherdrugs and/or therapies generally used for this conditions), duration,and/or frequency (including, for example, delaying or increasing time topain in an individual). As is understood by those skilled in the art,individuals may vary in terms of their response to treatment, and, assuch, for example, a “method of reducing incidence of pain in anindividual” reflects administering the NGF antibodies or fragmentsdescribed herein alone or in association with another active such as anNSAID or opioid as described herein, based on a reasonable expectationthat such administration may likely cause such a reduction in incidencein that particular individual.

“Ameliorating” pain or one or more symptoms of pain means a lessening orimprovement of one or more symptoms of a pain as compared to notadministering an NGF antibody or fragment alone or in association withanother active agonist such as an NSAID or opioid.

“Ameliorating” also includes shortening or reduction in duration of asymptom.

“Palliating” pain or one or more symptoms of pain means lessening theextent of one or more undesirable clinical manifestations of pain in anindividual or population of individuals treated with an NGF antibody orfragment in accordance with the invention.

As used therein, “delaying” the development of pain means to defer,hinder, slow, retard, stabilize, and/or postpone progression of pain.This delay can be of varying lengths of time, depending on the historyof the disease and/or individuals being treated. As is evident to oneskilled in the art, a sufficient or significant delay can, in effect,encompass prevention, in that the individual does not develop pain. Amethod that “delays” development of the symptom is a method that reducesprobability of developing the symptom in a given time frame and/orreduces extent of the symptoms in a given time frame, when compared tonot using the method. Such comparisons are typically based on clinicalstudies, using a statistically significant number of subjects.

For example, palliating” bone cancer pain (such as cancer painassociated with bone metastasis) or one or more symptoms of a bonecancer pain means lessening the extent of one or more undesirableclinical manifestations of bone cancer pain in an individual orpopulation of individuals treated with an NGF antibody or fragment inaccordance with the invention.

As used therein, “delaying” the development of pain, e.g., bone cancerpain including cancer pain associated with bone metastasis means todefer, hinder, slow, retard, stabilize, and/or postpone progression ofbone cancer pain including cancer pain associated with bone metastasis.This delay can be of varying lengths of time, depending on the historyof the disease and/or individuals being treated. As is evident to oneskilled in the art, a sufficient or significant delay can, in effect,encompass prevention, in that the individual does not develop bonecancer pain including cancer pain associated with bone metastasis. Amethod that “delays” development of the symptom is a method that reducesprobability of developing the symptom in a given time frame and/orreduces extent of the symptoms in a given time frame, when compared tonot using the method. Such comparisons are typically based on clinicalstudies, using a number of subjects sufficient to give a statisticallysignificant result.

“Development” or “progression” of pain, e.g., bone cancer pain includingcancer pain associated with bone metastasis means initial manifestationsand/or ensuing progression of the pain related disorder. Development ofbone cancer pain including cancer pain associated with bone metastasiscan be detectable and assessed using standard clinical techniques aswell known in the art. However, development also refers to progressionthat may be undetectable. For purpose of this invention, development orprogression refers to the biological course of the symptoms.

“Development” includes occurrence, recurrence, and onset. As used herein“onset” or “occurrence” of pain, e.g., bone cancer pain (such as cancerpain associated with bone metastasis) includes initial onset and/orrecurrence.

As used herein, “co-administration” includes simultaneous administrationand/or administration at different times. Co-administration alsoencompasses administration as a co-formulation (i.e., the NGF antibodyor fragment and another agent are present in the same composition) oradministration as separate compositions. As used herein,co-administration is meant to encompass any circumstance wherein anagent and NGF antibody or fragment are administered to an individual,which can occur simultaneously and/or separately. As further discussedherein, it is understood that the NGF antibody or fragment and an agentcan be administered at different dosing frequencies or intervals. Forexample, an anti-NGF antibody can be administered weekly, while theagent can be administered more frequently. It is understood that the NGFantagonist and the agent can be administered using the same route ofadministration or different routes of administration. Preferably theseanti-NGF antibody or fragment is one that

The term “opioid analgesic” refers to all drugs, natural or synthetic,with morphine-like actions. The synthetic and semi-synthetic opioidanalgesics are derivatives of five chemical classes of compound:phenanthrenes; phenylheptylamines; phenylpiperidines; morphinans; andbenzomorphans, all of which are within the scope of the term. Exemplaryopioid analgesics include codeine, dihydrocodeine, diacetylmorphine,hydrocodone, hydromorphone, levorphanol, oxymorphone, alfentanil,buprenorphine, butorphanol, fentanyl, sufentanyl, meperidine, methadone,nalbuphine, propoxyphene and pentazocine or pharmaceutically acceptablesalts thereof.

The term “NSAID” refers to a non-steroidal anti-inflammatory compound.NSAIDs are categorized by virtue of their ability to inhibitcyclooxygenase. Cyclooxygenase 1 and cyclooxygenase 2 are two majorisoforms of cyclooxygenase and most standard NSAIDs are mixed inhibitorsof the two isoforms. Most standard NSAIDs fall within one of thefollowing five structural categories: (1) propionic acid derivatives,such as ibuprofen, naproxen, naprosyn, diclofenac, and ketoprofen; (2)acetic acid derivatives, such as tolmetin and slindac; (3) fenamic acidderivatives, such as mefenamic acid and meclofenamic acid; (4)biphenylcarboxylic acid derivatives, such as diflunisal and flufenisal;and (5) oxicams, such as piroxim, sudoxicam, and isoxicam. Another classof NSAID has been described which selectively inhibit cyclooxygenase 2.Cox-2 inhibitors have been described, e.g., in U.S. Pat. Nos. 5,616,601;5,604,260; 5,593,994; 5,550,142; 5,536,752; 5,521,213; 5,475,995;5,639,780; 5,604,253; 5,552,422; 5,510,368; 5,436,265; 5,409,944; and5,130,311, all of which are hereby incorporated by reference. Certainexemplary COX-2 inhibitors include celecoxib (SC-58635), DUP-697,flosulide (CGP-28238), meloxicam, 6-methoxy-2 naphthylacetic acid(6-MNA), rofecoxib, MK-966, nabumetone (prodrug for 6-MNA), nimesulide,NS-398, SC-5766, SC-58215, T-614; or combinations thereof.

In some embodiments, aspirin and/or acetaminophen are taken inconjunction with NGF antibody or fragment. Aspirin is another type ofnon-steroidal anti-inflammatory compound.

Host Cell: In the present invention this is generally intended toinclude any cell that provides for the expression of antibodies orantibody fragments according to the invention. This includes by way ofexample bacterial, yeast, fungi, avian, plant cell, mammalian, andinsect cell expression systems. Typically antibodies are expressed inmammalian, bacterial and yeast cells. In a preferred embodiment thesubject antibodies or antibody fragments are expressed in a proprietarysecretory expression system that uses diploid Pichia yeast cultures forantibody expression. This expression system is disclosed in U.S. Pat.No. 7,927,863, by Cregg, issued Apr. 19, 2011, the contents of which areincorporated by reference herein.

Transgenic Animal or Plant: In the present invention this refers to anyanimal (non-human) or plant such as tobacco that has been geneticallymodified, e.g., by mutation of an endogenous gene, gene knock-in, geneknock-out, and the like. As is well known in the art transgenic animals,e.g., rodents, bovines, et al. and plants can be engineered with humanimmunoglobulin genes and thereby express human antibodies. Accordinglytransgenic animals and plants includes non-human animals and plantsengineered to express anti-NGF antibodies.

Mating competent yeast species: This is intended to broadly encompassany diploid or tetraploid yeast which can be grown in culture. Suchspecies of yeast may exist in a haploid, diploid, or other polyploidform. The cells of a given ploidy may, under appropriate conditions,proliferate for an indefinite number of generations in that form.Diploid cells can also sporulate to form haploid cells. Sequentialmating can result in tetraploid strains through further mating or fusionof diploid strains.

In a preferred embodiment of the invention, a mating competent yeast(Pichia) is used for expression. In a further preferred embodiment ofthe invention, the mating competent yeast of the genus Pichia is one ofthe following species: Pichia pastoris, Pichia methanolica, andHansenula polymorpha (Pichia angusta). In a particularly preferredembodiment of the invention, the mating competent yeast of the genusPichia is the species Pichia pastoris. However, as noted other hostcells may be used such as mammalian, insect, bacterial, fungal, as wellas non-human transgenic animals.

Selectable Marker: A selectable marker is a gene or gene fragment thatconfers a growth phenotype (physical growth characteristic) on a cellreceiving that gene as, for example through a transformation event. Theselectable marker allows that cell to survive and grow in a selectivegrowth medium under conditions in which cells that do not receive thatselectable marker gene cannot grow. Selectable marker genes generallyfall into several types, including positive selectable marker genes suchas a gene that confers on a cell resistance to an antibiotic or otherdrug, temperature when two ts mutants are crossed or a ts mutant istransformed; negative selectable marker genes such as a biosyntheticgene that confers on a cell the ability to grow in a medium without aspecific nutrient needed by all cells that do not have that biosyntheticgene, or a mutagenized biosynthetic gene that confers on a cellinability to grow by cells that do not have the wild type gene; and thelike. Suitable markers include but are not limited to: ZEO; G418; LYS3;MET1; MET3a; ADE1; ADE3; URA3; and the like.

Expression Vector: These DNA vectors contain elements that facilitatemanipulation for the expression of a foreign protein within the targethost cell. Conveniently, manipulation of sequences and production of DNAfor transformation is first performed in a bacterial host, e.g. E. coli,and usually vectors will include sequences to facilitate suchmanipulations, including a bacterial origin of replication andappropriate bacterial selection marker. Selection markers encodeproteins necessary for the survival or growth of transformed host cellsgrown in a selective culture medium. Host cells not transformed with thevector containing the selection gene will not survive in the culturemedium. Typical selection genes encode proteins that (a) conferresistance to antibiotics or other toxins, (b) complement auxotrophicdeficiencies, or (c) supply critical nutrients not available fromcomplex media. Exemplary vectors and methods for transformation of yeastare described, for example, in Burke, D., Dawson, D., & Stearns, T.(2000). Methods in yeast genetics: a Cold Spring Harbor Laboratorycourse manual. Plainview, N.Y.: Cold Spring Harbor Laboratory Press.

Expression vectors for use in the methods of the invention willgenerally further include host cell specific sequences, including aselectable auxotrophic or drug marker for identifying transformed hostcells. A drug marker may further be used to amplify copy number of thevector in a host cell.

The antibody polypeptide coding sequence of interest is operably linkedto transcriptional and translational regulatory sequences that providefor expression of the polypeptide in host cells. These vector componentsmay include, but are not limited to, one or more of the following: anenhancer element, a promoter, and a transcription termination sequence.Sequences for the secretion of the polypeptide may also be included,e.g. a signal sequence, and the like. A yeast origin of replication isoptional, as expression vectors are often integrated into the host cellgenome. In one embodiment of the invention, the polypeptide of interestis operably linked, or fused, to sequences providing for optimizedsecretion of the polypeptide from host cells.

Nucleic acids are “operably linked” when placed into a functionalrelationship with another nucleic acid sequence. For example, DNA for asignal sequence is operably linked to DNA for a polypeptide if it isexpressed as a preprotein that participates in the secretion of thepolypeptide; a promoter or enhancer is operably linked to a codingsequence if it affects the transcription of the sequence. Generally,“operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading frame. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites oralternatively via a PCR/recombination method familiar to those skilledin the art (GatewayR Technology; Invitrogen, Carlsbad Calif.). If suchsites do not exist, the synthetic oligonucleotide adapters or linkersare used in accordance with conventional practice.

Promoters are untranslated sequences located upstream (5′) to the startcodon of a structural gene (generally within about 100 to 1000 bp) thatcontrol the transcription and translation of particular nucleic acidsequences to which they are operably linked. Such promoters fall intoseveral classes: inducible, constitutive, and repressible promoters(that increase levels of transcription in response to absence of arepressor). Inducible promoters may initiate increased levels oftranscription from DNA under their control in response to some change inculture conditions, e.g., the presence or absence of a nutrient or achange in temperature.

The promoter fragment may also serve as the site for homologousrecombination and integration of the expression vector into the samesite in the host cell genome; alternatively a selectable marker is usedas the site for homologous recombination.

The antibody polypeptides of interest may be produced recombinantly notonly directly, but also as a fusion polypeptide with a heterologouspolypeptide, e.g. a signal sequence or other polypeptide having aspecific cleavage site at the N-terminus of the mature protein orpolypeptide. In general, the signal sequence may be a component of thevector, or it may be a part of the polypeptide coding sequence that isinserted into the vector. The heterologous signal sequence selectedpreferably is one that is recognized and processed through one of thestandard pathways available within the host cell. Secretion signals ofinterest include mammalian, yeast, and bacterial signal sequences, whichmay be endogenous or heterologous to the host cell or antibody proteinbeing secreted, or may be a native sequence for the antibody proteinbeing secreted. Signal sequences include pre-peptide sequences, and insome instances may include propeptide sequences. Many such signalsequences are known in the art, including the signal sequences found onimmunoglobulin chains, e.g., K28 preprotoxin sequence, PHA-E, FACE,human MCP-1, human serum albumin signal sequences, human Ig heavy chain,human Ig light chain, and the like. For example, see Hashimoto et. al.Protein Eng 11(2) 75 (1998); and Kobayashi et. al. Therapeutic Apheresis2(4) 257 (1998).

Transcription may be increased by inserting a transcriptional activatorsequence into the vector. These activators are cis-acting elements ofDNA, usually about from 10 to 300 bp, which act on a promoter toincrease its transcription. Transcriptional enhancers are relativelyorientation and position independent, having been found 5′ and 3′ to thetranscription unit, within an intron, as well as within the codingsequence itself. The enhancer may be spliced into the expression vectorat a position 5′ or 3′ to the coding sequence, but is preferably locatedat a site 5′ from the promoter.

Expression vectors used in eukaryotic host cells may also containsequences necessary for the termination of transcription and forstabilizing the mRNA. Such sequences are commonly available from 3′ tothe translation termination codon, in untranslated regions of eukaryoticor viral DNAs or cDNAs. These regions contain nucleotide segmentstranscribed as polyadenylated fragments in the untranslated portion ofthe mRNA.

Construction of suitable vectors containing one or more of theabove-listed components employs standard ligation techniques orPCR/recombination methods. Isolated plasmids or DNA fragments arecleaved, tailored, and re-ligated in the form desired to generate theplasmids required or via recombination methods. For analysis to confirmcorrect sequences in plasmids constructed, the ligation mixtures areused to transform host cells, and successful transformants selected byantibiotic resistance (e.g. ampicillin or Zeocin) where appropriate.Plasmids from the transformants are prepared, analyzed by restrictionendonuclease digestion and/or sequenced.

As an alternative to restriction and ligation of fragments,recombination methods based on att sites and recombination enzymes maybe used to insert DNA sequences into a vector. Such methods aredescribed, for example, by Landy (1989) Ann. Rev. Biochem. 58:913-949;and are known to those of skill in the art. Such methods utilizeintermolecular DNA recombination that is mediated by a mixture of lambdaand E. coli-encoded recombination proteins. Recombination occurs betweenspecific attachment (att) sites on the interacting DNA molecules. For adescription of att sites see Weisberg and Landy (1983) Site-SpecificRecombination in Phage Lambda, in Lambda II, Weisberg, ed. (Cold SpringHarbor, N.Y.: Cold Spring Harbor Press), pp. 211-250. The DNA segmentsflanking the recombination sites are switched, such that afterrecombination, the att sites are hybrid sequences comprised of sequencesdonated by each parental vector. The recombination can occur betweenDNAs of any topology.

Att sites may be introduced into a sequence of interest by ligating thesequence of interest into an appropriate vector; generating a PCRproduct containing att B sites through the use of specific primers;generating a cDNA library cloned into an appropriate vector containingatt sites; and the like.

Folding, as used herein, refers to the three-dimensional structure ofpolypeptides and proteins, where interactions between amino acidresidues act to stabilize the structure. While non-covalent interactionsare important in determining structure, usually the proteins of interestwill have intra- and/or intermolecular covalent disulfide bonds formedby two cysteine residues. For naturally occurring proteins andpolypeptides or derivatives and variants thereof, the proper folding istypically the arrangement that results in optimal biological activity,and can conveniently be monitored by assays for activity, e.g. ligandbinding, enzymatic activity, etc.

In some instances, for example where the desired product is of syntheticorigin, assays based on biological activity will be less meaningful. Theproper folding of such molecules may be determined on the basis ofphysical properties, energetic considerations, modeling studies, and thelike.

The expression host may be further modified by the introduction ofsequences encoding one or more enzymes that enhance folding anddisulfide bond formation, i.e. foldases, chaperonins, etc. Suchsequences may be constitutively or inducibly expressed in the host cell,using vectors, markers, etc. as known in the art. Preferably thesequences, including transcriptional regulatory elements sufficient forthe desired pattern of expression, are stably integrated in the host orhost cell genome through a targeted methodology.

The terms “desired protein” or “desired antibody” are usedinterchangeably and refer generally to a parent antibody specific to atarget, i.e., NGF or a chimeric or humanized antibody or a bindingportion thereof derived therefrom as described herein. The term“antibody” is intended to include any polypeptide chain-containingmolecular structure with a specific shape that fits to and recognizes anepitope, where one or more non-covalent binding interactions stabilizethe complex between the molecular structure and the epitope. Thearchetypal antibody molecule is the immunoglobulin, and all types ofimmunoglobulins, IgG, IgM, IgA, IgE, IgD, etc., from all sources, e.g.human, rodent, rabbit, cow, sheep, pig, dog, other mammals, chicken,other avians, etc., are considered to be “antibodies.” A preferredsource for producing antibodies useful as starting material according tothe invention is rabbits. Numerous antibody coding sequences have beendescribed; and others may be raised by methods well-known in the art.Examples thereof include chimeric antibodies, human antibodies and othernon-human mammalian antibodies, humanized antibodies, single chainantibodies (such as scFvs), camelbodies, nanobodies, MetMab likemonovalent antibody fragments, IgNAR (single-chain antibodies derivedfrom sharks), small-modular immunopharmaceuticals (SMIPs), and antibodyfragments such as Fabs, Fab′, and the like. See Streltsov V A, et al.,Structure of a shark IgNAR antibody variable domain and modeling of anearly-developmental isotype, Protein Sci. 2005 November; 14(11):2901-9.Epub 2005 Sep. 30; Greenberg A S, et al., A new antigen receptor genefamily that undergoes rearrangement and extensive somaticdiversification in sharks, Nature. 1995 Mar. 9; 374(6518):168-73;Nuttall S D, et al., Isolation of the new antigen receptor fromwobbegong sharks, and use as a scaffold for the display of protein looplibraries, Mol Immunol. 2001 August; 38(4):313-26; Hamers-Casterman C,et al., Naturally occurring antibodies devoid of light chains, Nature.1993 Jun. 3; 363(6428):446-8; Gill D S, et al., Biopharmaceutical drugdiscovery using novel protein scaffolds, Curr Opin Biotechnol. 2006December; 17(6):653-8. Epub 2006 Oct. 19. The present invention includesin particular monovalent antibody molecules that bind NGF, which areanalogous to MetMab molecules. MetMab is a monovalent antibody specificto Met. (“Met” refers to a protein encoded by the nucleotide sequenceset forth in Park et al., Proc. Natl. Acad. Sci. 84, 7479-(1987), orfragments thereof, as well as related polypeptides, which include, butare not limited to, allelic variants, splice variants, derivativevariants, substitution variants, deletion variants, and/or insertionvariants, fusion polypeptides, and interspecies homologs). The MetMabantibody, is a monovalent antibody known by different names includingOA-5d5 (Genentech) (also called One Armed 5d5, 5d5, MetMab, PRO143966,among others). Antibody OA-5d5, including its structure and properties,and methods for making and using it, are described in U.S. PublicationNo. 2007/0092520. In one embodiment, an anti-NGF antibody according tothe invention may comprise a single Fab region linked to an Fc region.In such embodiment, an antibody of the invention may comprise light andheavy chain variable domains as described herein. In such an embodiment,the antibody is monovalent and may comprise an intact Fc region. Inanother such embodiment, the Fc region may comprise at least oneprotuberance (knob) and at least one cavity (hole), wherein the presenceof the protuberance and cavity enhances formation of a complex betweenan Fc polypeptide comprising the protuberance and an Fc polypeptidecomprising the cavity, for example as described in WO 2005/063816. Inone embodiment, the Fc region of an antibody of the invention maycomprise a first and a second Fc polypeptide, wherein the first andsecond polypeptide each comprises one or more mutations with respect towild type human Fc. In one embodiment, a cavity mutation is T366S, L368Aand/or Y407V. In another embodiment, a protuberance mutation is T366W.In a specific embodiment, a monovalent antibody according to the subjectinvention may comprise a one-armed antibody synthesized as described inWO2005/063816. In such embodiment, the one-armed antibody may compriseFc mutations constituting “knobs” and “holes” as described inWO2005/063816. For example, a hole mutation can be one or more of T366A,L368A and/or Y407V in an Fc polypeptide, and a cavity mutation can beT366W.

Antibodies or antigen binding fragments are typically produced bygenetic engineering. In this technique, as with other methods,antibody-producing cells are sensitized to the desired antigen orimmunogen. The messenger RNA isolated from antibody producing cells isused as a template to make cDNA using PCR amplification. A library ofvectors, each containing one heavy chain gene and one light chain generetaining the initial antigen specificity, is produced by insertion ofappropriate sections of the amplified immunoglobulin cDNA into theexpression vectors. A combinatorial library is constructed by combiningthe heavy chain gene library with the light chain gene library. Thisresults in a library of clones which co-express a heavy and light chain(resembling the Fab fragment or antigen binding fragment of an antibodymolecule). The vectors that carry these genes are co-transfected into ahost cell. When antibody gene synthesis is induced in the transfectedhost, the heavy and light chain proteins self-assemble to produce activeantibodies that can be detected by screening with the antigen orimmunogen.

Antibody coding sequences of interest include those encoded by nativesequences, as well as nucleic acids that, by virtue of the degeneracy ofthe genetic code, are not identical in sequence to the disclosed nucleicacids, and variants thereof. Variant polypeptides can include amino acid(aa) substitutions, additions or deletions. The amino acid substitutionscan be conservative amino acid substitutions or substitutions toeliminate non-essential amino acids, such as to alter a glycosylationsite, or to minimize misfolding by substitution or deletion of one ormore cysteine residues that are not necessary for function. Variants canbe designed so as to retain or have enhanced biological activity of aparticular region of the protein (e.g., a functional domain, catalyticamino acid residues, etc). Variants also include fragments of thepolypeptides disclosed herein, particularly biologically activefragments and/or fragments corresponding to functional domains.Techniques for in vitro mutagenesis of cloned genes are known. Alsoincluded in the subject invention are polypeptides that have beenmodified using ordinary molecular biological techniques so as to improvetheir resistance to proteolytic degradation or to optimize solubilityproperties or to render them more suitable as a therapeutic agent.

Chimeric antibodies herein include any antibody or antibody fragmentthat is obtained by combining portions or fragments of least 2antibodies, which may be of the same or different species. Chimericantibodies include humanized antibodies which comprise amino acidsresidues of human and non-human origin, most typically most or all ofthe CDRs of a rodent or rabbit antibody and most or all of the frameworkresidues from a human antibody. Chimeric antibodies may be made byrecombinant means, e.g., by combining the variable light and heavy chainregions (V_(L) and V_(H)), obtained from antibody producing cells of onespecies with the constant light and heavy chain regions from another.Typically chimeric antibodies utilize rodent or rabbit variable regionsand human constant regions, in order to produce an antibody withpredominantly human domains. The production of chimeric antibodies iswell known in the art, and may be achieved by well known methods (e.g.,as described, e.g., in U.S. Pat. No. 5,624,659, incorporated herein byreference in its entirety). Chimeric antibodies may comprise differentconstant domains, especially human or non-human primate constantdomains. It is specifically contemplated that the constant regionscontained in the chimeric antibodies of the invention may be selectedfrom human IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10,IgG11, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 or IgG19 constantregions or may comprise non-human primate constant regions.

As noted, humanized antibodies, which are a type of chimeric antibody,are typically engineered to contain human-like immunoglobulin domains,and may incorporate only complementarity-determining regions (all ormost) of the animal-derived antibody and typically comprise few or evenno animal-derived framework residues. This is accomplished by carefullyexamining the sequence of the hyper-variable loops of the variableregions of the monoclonal antibody, and fitting them to the structure ofthe human antibody chains. Although facially complex, the process isstraightforward in practice. See, e.g., U.S. Pat. No. 6,187,287,incorporated fully herein by reference.

In addition to entire immunoglobulins (or their recombinantcounterparts), immunoglobulin fragments comprising the epitope bindingsite (e.g., Fab′, F(ab′)₂, or other fragments) may be synthesized.“Fragment,” or minimal immunoglobulins may be designed utilizingrecombinant immunoglobulin techniques. For instance “Fv” immunoglobulinsfor use in the present invention may be produced by synthesizing a fusedvariable light chain region and a variable heavy chain region.Combinations of antibodies are also of interest, e.g. diabodies, whichcomprise two distinct Fv specificities. In another embodiment of theinvention, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, MetMab like monovalent antibodies and IgNAR are encompassedby immunoglobulin fragments.

Immunoglobulins and fragments thereof may be modifiedpost-translationally, e.g. to add effector moieties such as chemicallinkers, detectable moieties, such as fluorescent dyes, enzymes, toxins,substrates, bioluminescent materials, radioactive materials,chemiluminescent moieties and the like, or specific binding moieties,such as streptavidin, avidin, or biotin, and the like may be utilized inthe methods and compositions of the present invention. Examples ofadditional effector molecules are provided infra.

A polynucleotide sequence “corresponds” to a polypeptide sequence iftranslation of the polynucleotide sequence in accordance with thegenetic code yields the polypeptide sequence (i.e., the polynucleotidesequence “encodes” the polypeptide sequence), one polynucleotidesequence “corresponds” to another polynucleotide sequence if the twosequences encode the same polypeptide sequence.

A “heterologous” region or domain of a DNA construct is an identifiablesegment of DNA within a larger DNA molecule that is not found inassociation with the larger molecule in nature. Thus, when theheterologous region encodes a mammalian gene, the gene will usually beflanked by DNA that does not flank the mammalian genomic DNA in thegenome of the source organism. Another example of a heterologous regionis a construct where the coding sequence itself is not found in nature(e.g., a cDNA where the genomic coding sequence contains introns, orsynthetic sequences having codons different than the native gene).Allelic variations or naturally-occurring mutational events do not giverise to a heterologous region of DNA as defined herein.

A “coding sequence” is an in-frame sequence of codons that (in view ofthe genetic code) correspond to or encode a protein or peptide sequence.Two coding sequences correspond to each other if the sequences or theircomplementary sequences encode the same amino acid sequences. A codingsequence in association with appropriate regulatory sequences may betranscribed and translated into a polypeptide. A polyadenylation signaland transcription termination sequence will usually be located 3′ to thecoding sequence. A “promoter sequence” is a DNA regulatory regioncapable of binding RNA polymerase in a cell and initiating transcriptionof a downstream (3′ direction) coding sequence. Promoter sequencestypically contain additional sites for binding of regulatory molecules(e.g., transcription factors) which affect the transcription of thecoding sequence. A coding sequence is “under the control” of thepromoter sequence or “operatively linked” to the promoter when RNApolymerase binds the promoter sequence in a cell and transcribes thecoding sequence into mRNA, which is then in turn translated into theprotein encoded by the coding sequence.

Vectors are used to introduce a foreign substance, such as DNA, RNA orprotein, into an organism or host cell. Typical vectors includerecombinant viruses (for polynucleotides) and liposomes (forpolypeptides). A “DNA vector” is a replicon, such as plasmid, phage orcosmid, to which another polynucleotide segment may be attached so as tobring about the replication of the attached segment. An “expressionvector” is a DNA vector which contains regulatory sequences which willdirect polypeptide synthesis by an appropriate host cell. This usuallymeans a promoter to bind RNA polymerase and initiate transcription ofmRNA, as well as ribosome binding sites and initiation signals to directtranslation of the mRNA into a polypeptide(s). Incorporation of apolynucleotide sequence into an expression vector at the proper site andin correct reading frame, followed by transformation of an appropriatehost cell by the vector, enables the production of a polypeptide encodedby said polynucleotide sequence.

“Amplification” of polynucleotide sequences is the in vitro productionof multiple copies of a particular nucleic acid sequence. The amplifiedsequence is usually in the form of DNA. A variety of techniques forcarrying out such amplification are described in a review article by VanBrunt (1990, Bio/Technol., 8(4):291-294). Polymerase chain reaction orPCR is a prototype of nucleic acid amplification, and use of PCR hereinshould be considered exemplary of other suitable amplificationtechniques.

The general structure of antibodies in vertebrates now is wellunderstood (Edelman, G. M., Ann. N.Y. Acad. Sci., 190: 5 (1971)).Antibodies consist of two identical light polypeptide chains ofmolecular weight approximately 23,000 daltons (the “light chain”), andtwo identical heavy chains of molecular weight 53,000-70,000 (the “heavychain”). The four chains are joined by disulfide bonds in a “Y”configuration wherein the light chains bracket the heavy chains startingat the mouth of the “Y” configuration. The “branch” portion of the “Y”configuration is designated the Fab region; the stem portion of the “Y”configuration is designated the Fc region. The amino acid sequenceorientation runs from the N-terminal end at the top of the “Y”configuration to the C-terminal end at the bottom of each chain. TheN-terminal end possesses the variable region having specificity for theantigen that elicited it, and is approximately 100 amino acids inlength, there being slight variations between light and heavy chain andfrom antibody to antibody.

The variable region is linked in each chain to a constant region thatextends the remaining length of the chain and that within a particularclass of antibody does not vary with the specificity of the antibody(i.e., the antigen eliciting it). There are five known major classes ofconstant regions that determine the class of the immunoglobulin molecule(IgG, IgM, IgA, IgD, and IgE corresponding to γ, μ, α, δ, and ε (gamma,mu, alpha, delta, or epsilon) heavy chain constant regions). Theconstant region or class determines subsequent effector function of theantibody, including activation of complement (Kabat, E. A., StructuralConcepts in Immunology and Immunochemistry, 2nd Ed., p. 413-436, Holt,Rinehart, Winston (1976)), and other cellular responses (Andrews, D. W.,et al., Clinical Immunobiology, pp 1-18, W. B. Sanders (1980); Kohl, S.,et al., Immunology, 48: 187 (1983)); while the variable regiondetermines the antigen with which it will react. Light chains areclassified as either κ (kappa) or λ (lambda). Each heavy chain class canbe prepared with either kappa or lambda light chain. The light and heavychains are covalently bonded to each other, and the “tail” portions ofthe two heavy chains are bonded to each other by covalent disulfidelinkages when the immunoglobulins are generated either by hybridomas orby B cells.

The expression “variable region” or “VR” refers to the domains withineach pair of light and heavy chains in an antibody that are involveddirectly in binding the antibody to the antigen. Each heavy chain has atone end a variable domain (V_(H)) followed by a number of constantdomains. Each light chain has a variable domain (VL) at one end and aconstant domain at its other end; the constant domain of the light chainis aligned with the first constant domain of the heavy chain, and thelight chain variable domain is aligned with the variable domain of theheavy chain.

The expressions “complementarity determining region,” “hypervariableregion,” or “CDR” refer to one or more of the hyper-variable orcomplementarity determining regions (CDRs) found in the variable regionsof light or heavy chains of an antibody (See Kabat, E. A. et al.,Sequences of Proteins of Immunological Interest, National Institutes ofHealth, Bethesda, Md., (1987)). These expressions include thehypervariable regions as defined by Kabat et al. (“Sequences of Proteinsof Immunological Interest,” Kabat E., et al., US Dept. of Health andHuman Services, 1983) or the hypervariable loops in 3-dimensionalstructures of antibodies (Chothia and Lesk, J Mol. Biol. 196 901-917(1987)). The CDRs in each chain are held in close proximity by frameworkregions and, with the CDRs from the other chain, contribute to theformation of the antigen binding site. Within the CDRs there are selectamino acids that have been described as the selectivity determiningregions (SDRs) which represent the critical contact residues used by theCDR in the antibody-antigen interaction (Kashmiri, S., Methods, 36:25-34(2005)).

The expressions “framework region” or “FR” refer to one or more of theframework regions within the variable regions of the light and heavychains of an antibody (See Kabat, E. A. et al., Sequences of Proteins ofImmunological Interest, National Institutes of Health, Bethesda, Md.,(1987)). These expressions include those amino acid sequence regionsinterposed between the CDRs within the variable regions of the light andheavy chains of an antibody.

Anti-NGF Antibodies and Binding Fragments Thereof Having BindingActivity for NGF Antibody Ab1

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is antibody Ab1 or fragments thereof, for example as setforth below, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA and the association of NGF with p75. In oneembodiment, the invention includes chimeric antibodies having bindingspecificity to NGF and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 1) ALVMTQTPSSVSAAVGGTVTINCQASQNIYSNLAWYQQRPGQRPKLLIYGASNLDAGVPSRFRGSGSGTEYTLTISDLECDDVGTYYCQSAFDSDSTENT FGGGTEVVVKR.

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF andpossessing a light chain sequence comprising the sequence set forthbelow:

(SEQ ID NO: 2) ALVMTQTPSSVSAAVGGTVTINCQASQNIYSNLAWYQQRPGQRPKLLIYGASNLDAGVPSRFRGSGSGTEYTLTISDLECDDVGTYYCQSAFDSDSTENTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF andpossessing a variable heavy chain sequence comprising the sequence setforth below:

(SEQ ID NO: 3) QSLEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGVITSIGSTVYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARGYDDYD EMTYFNIWGQGTLVTVSS.

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF andpossessing a heavy chain sequence comprising the sequence set forthbelow:

(SEQ ID NO: 4) QSLEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGVITSIGSTVYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARGYDDYDEMTYFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiescomprise one or more of the polypeptide sequences of SEQ ID NO: 5; SEQID NO: 6; and SEQ ID NO: 7 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 1 or the light chainsequence of SEQ ID NO: 2, and/or one or more of the polypeptidesequences of SEQ ID NO: 8; SEQ ID NO: 9; and SEQ ID NO: 10 whichcorrespond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable heavy chain sequence of SEQ IDNO: 3 or the heavy chain sequence of SEQ ID NO: 4, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the CDRs, thevariable heavy and variable light chain sequences, and the heavy andlight chain sequences set forth above, including all of them.

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibody is afragment having binding specificity to NGF. In one embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 1 orSEQ ID NO: 2. In another embodiment of the invention, antibody fragmentsof the invention comprise, or alternatively consist of, the polypeptidesequence of SEQ ID NO: 3 or SEQ ID NO: 4.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of paincomprise, or alternatively consist of, one or more of the polypeptidesequences of SEQ ID NO: 5; SEQ ID NO: 6; and SEQ ID NO: 7 whichcorrespond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 1 or the light chain sequence of SEQ ID NO: 2.

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude fragments having binding specificity to NGF comprise, oralternatively consist of, one or more of the polypeptide sequences ofSEQ ID NO: 8; SEQ ID NO: 9; and SEQ ID NO: 10 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 3 or the heavy chainsequence of SEQ ID NO: 4.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 1; the variable heavy chain region of SEQ IDNO: 3; the complementarity-determining regions (SEQ ID NO: 5; SEQ ID NO:6; and SEQ ID NO: 7) of the variable light chain region of SEQ ID NO: 1;and the complementarity-determining regions (SEQ ID NO: 8; SEQ ID NO: 9;and SEQ ID NO: 10) of the variable heavy chain region of SEQ ID NO: 3.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody is Ab1, comprising, or alternatively consisting of,SEQ ID NO: 2 and SEQ ID NO: 4, and having at least one of the biologicalactivities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for use herein comprise, or alternatively consist ofFab (fragment antigen binding) fragments having binding specificity forNGF or may comprise MetMab like monovalent antibodies. With respect toantibody Ab1, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 1 and the variable heavy chain sequence of SEQ IDNO: 3. This embodiment of the invention further contemplates additions,deletions, and variants of SEQ ID NO: 1 and/or SEQ ID NO: 3 in said Fabwhile retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab1.In another embodiment of the invention, anti-NGF antibodies such as Ab1or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, plant cell, plant,animal, insect, or microbial systems such as yeast cells (for examplediploid yeast such as diploid Pichia) and other yeast strains. SuitablePichia species include, but are not limited to, Pichia pastoris.

Antibody Ab2

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is antibody Ab2 or fragments thereof, for example as setforth below, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA and the association of NGF with p75. In oneembodiment, the invention includes chimeric or humanized antibodieshaving binding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 11) DIQMTQSPSTLSASVGDRVTITCQASQNIYSNLAWYQQKPGKAPKLLIYGASNLDAGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQSAFDSDSTENT FGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 12) DIQMTQSPSTLSASVGDRVTITCQASQNIYSNLAWYQQKPGKAPKLLIYGASNLDAGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQSAFDSDSTENTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies havingbinding specificity to NGF for treatment or prevention of pain and painassociated conditions and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 13) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYAMSWVRQAPGKGLEWVGVITSIGSTVYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYDDYDEMTYFNIWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies havingbinding specificity to NGF for treatment or prevention of pain and painassociated conditions and possessing a heavy chain sequence comprisingthe sequence set forth below:

(SEQ ID NO: 14) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYAMSWVRQAPGKGLEWVGVITSIGSTVYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYDDYDEMTYFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies or antibody fragments fortreatment or prevention of pain and pain associated conditionscomprising one or more of the polypeptide sequences of SEQ ID NO: 15;SEQ ID NO: 16; and SEQ ID NO: 17 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 11 or the light chainsequence of SEQ ID NO: 12, and/or one or more of the polypeptidesequences of SEQ ID NO: 18; SEQ ID NO: 19; and SEQ ID NO: 20 whichcorrespond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable heavy chain sequence of SEQ IDNO: 13 or the heavy chain sequence of SEQ ID NO: 14, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the CDRs, thevariable heavy and variable light chain sequences, and the heavy andlight chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 11 or SEQ ID NO: 12. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 13 or SEQ ID NO: 14.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 15; SEQ ID NO:16; and SEQ ID NO: 17 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 11 or the light chainsequence of SEQ ID NO: 12.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 18; SEQ ID NO:19; and SEQ ID NO: 20 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 13 or the heavy chainsequence of SEQ ID NO: 14.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF for treatment or prevention of pain and pain associated conditionscomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 11; the variable heavy chain region of SEQ IDNO: 13; the complementarity-determining regions (SEQ ID NO: 15; SEQ IDNO: 16; and SEQ ID NO: 17) of the variable light chain region of SEQ IDNO: 11; and the complementarity-determining regions (SEQ ID NO: 18; SEQID NO: 19; and SEQ ID NO: 20) of the variable heavy chain region of SEQID NO: 13.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab2, comprising, or alternatively consistingof, SEQ ID NO: 12 and SEQ ID NO: 14, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFand also MetMab like monovalent antibodies. With respect to antibodyAb2, the Fab fragment includes the variable light chain sequence of SEQID NO: 11 and the variable heavy chain sequence of SEQ ID NO: 13. Thisembodiment of the invention further contemplates additions, deletions,and variants of SEQ ID NO: 11 and/or SEQ ID NO: 13 in said Fab whileretaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab2.In another embodiment of the invention, anti-NGF antibodies such as Ab2or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,plant, animal, or microbial systems such as bacterial or yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab3

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is Ab3 or fragments thereof, for example as set forthbelow, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA without appreciably inhibiting theassociation of NGF with p75. In one embodiment, the invention includeschimeric antibodies having binding specificity to NGF and possessing avariable light chain sequence comprising the sequence set forth below:

(SEQ ID NO: 21) AVLTQTPSPVSAAMGDTVTIKCQSSQSVYKNNYLSWYQQKPGQPPRLLIYDASNLPSGVPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGDYDDDADNA FGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 22) AVLTQTPSPVSAAMGDTVTIKCQSSQSVYKNNYLSWYQQKPGQPPRLLIYDASNLPSGVPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGDYDDDADNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 23) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYVMIWVRQAPGKGLEYIGITWSAGTYYASWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCAGGGGSIY DIWGPGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 24) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYVMIWVRQAPGKGLEYIGITWSAGTYYASWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCAGGGGSIYDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies or antibody fragments fortreatment or prevention of pain and pain associated conditionscomprising one or more of the polypeptide sequences of SEQ ID NO: 25;SEQ ID NO: 26; and SEQ ID NO: 27 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 21 or the light chainsequence of SEQ ID NO: 22, and/or one or more of the polypeptidesequences of SEQ ID NO: 28; SEQ ID NO: 29; and SEQ ID NO: 30 whichcorrespond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable heavy chain sequence of SEQ IDNO: 23 or the heavy chain sequence of SEQ ID NO: 24, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the CDRs, thevariable heavy and variable light chain sequences, and the heavy andlight chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 21 or SEQ ID NO: 22. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 23 or SEQ ID NO: 24.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 25; SEQ ID NO:26; and SEQ ID NO: 27 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 21 or the light chainsequence of SEQ ID NO: 22.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 28; SEQ ID NO:29; and SEQ ID NO: 30 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 23 or the heavy chainsequence of SEQ ID NO: 24.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein for treatment orprevention of pain and pain associated conditions. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 21; the variable heavy chain region of SEQ IDNO: 23; the complementarity-determining regions (SEQ ID NO: 25; SEQ IDNO: 26; and SEQ ID NO: 27) of the variable light chain region of SEQ IDNO: 21; and the complementarity-determining regions (SEQ ID NO: 28; SEQID NO: 29; and SEQ ID NO: 30) of the variable heavy chain region of SEQID NO: 23.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab3, comprising, or alternatively consistingof, SEQ ID NO: 22 and SEQ ID NO: 24, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab3, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 21 and the variable heavy chain sequence of SEQID NO: 23. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 21 and/or SEQ ID NO: 23in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab3.In another embodiment of the invention, anti-NGF antibodies such as Ab3or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, plant cells, transgenic plantsand animals, fungal, insect, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab4

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is antibody Ab4 or fragments thereof, for example as setforth below, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA without appreciably inhibiting theassociation of NGF with p75 and/or for preventing or effectivelytreating pain. In one embodiment, the invention includes chimeric orhumanized antibodies for treatment or prevention of pain and painassociated conditions having binding specificity to NGF and possessing avariable light chain sequence comprising the sequence set forth below:

(SEQ ID NO: 31) DIQMTQSPSTLSASVGDRVTITCQSSQSVYKNNYLSWYQQKPGKAPKLLIYDASNLPSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCLGDYDDDADN AFGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 32) DIQMTQSPSTLSASVGDRVTITCQSSQSVYKNNYLSWYQQKPGKAPKLLIYDASNLPSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCLGDYDDDADNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 33) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYVMIWVRQAPGKGLEYIGITWSAGTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGGGS IYDIWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 34) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYVMIWVRQAPGKGLEYIGITWSAGTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGGGSIYDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 35; SEQ ID NO: 36; and SEQ IDNO: 37 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 31 or the light chain sequence of SEQ ID NO: 32, and/or oneor more of the polypeptide sequences of SEQ ID NO: 38; SEQ ID NO: 39;and SEQ ID NO: 40 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 33 or the heavy chain sequence of SEQ ID NO: 34,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 31 or SEQ ID NO: 32. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 33 or SEQ ID NO: 34.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 35; SEQ ID NO:36; and SEQ ID NO: 37 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 31 or the light chainsequence of SEQ ID NO: 32.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 38; SEQ ID NO:39; and SEQ ID NO: 40 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 33 or the heavy chainsequence of SEQ ID NO: 34.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 31; the variable heavy chain region of SEQ IDNO: 33; the complementarity-determining regions (SEQ ID NO: 35; SEQ IDNO: 36; and SEQ ID NO: 37) of the variable light chain region of SEQ IDNO: 31; and the complementarity-determining regions (SEQ ID NO: 38; SEQID NO: 39; and SEQ ID NO: 40) of the variable heavy chain region of SEQID NO: 33.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab4, comprising, or alternatively consistingof, SEQ ID NO: 32 and SEQ ID NO: 34, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab4, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 31 and the variable heavy chain sequence of SEQID NO: 33. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 31 and/or SEQ ID NO: 33in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) of Ab4.In another embodiment of the invention, anti-NGF antibodies such as Ab4or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab5

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab5 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and the association of NGF with p75. In one embodiment, theinvention includes chimeric antibodies having binding specificity to NGFand possessing a variable light chain sequence comprising the sequenceset forth below:

(SEQ ID NO: 41) AYDMTQTPASVEVAVGGTVTIKCQASQSIYSNLAWYQQRPGQPPKLLIYDASTLESGVPSRFKGSGSGTEYTLTISGVECADAASYYCQQGFTVSDIDNA FGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 42) AYDMTQTPASVEVAVGGTVTIKCQASQSIYSNLAWYQQRPGQPPKLLIYDASTLESGVPSRFKGSGSGTEYTLTISGVECADAASYYCQQGFTVSDIDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 43) QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAVGWVRQAPGKGLEWIGIIGRNGNTWYASWARGRFTISKTSTTVDLKITSPTSEDTATYFCARGYGRSV AYYVFNIWGPGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 44) QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAVGWVRQAPGKGLEWIGIIGRNGNTWYASWARGRFTISKTSTTVDLKITSPTSEDTATYFCARGYGRSVAYYVFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 45; SEQ ID NO: 46; and SEQ IDNO: 47 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 41 or the light chain sequence of SEQ ID NO: 42, and/or oneor more of the polypeptide sequences of SEQ ID NO: 48; SEQ ID NO: 49;and SEQ ID NO: 50 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 43 or the heavy chain sequence of SEQ ID NO: 44,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 41 or SEQ ID NO: 42. In anotherembodiment of the invention, antibody fragments of the invention fortreatment or prevention of pain and pain associated conditions comprise,or alternatively consist of, the polypeptide sequence of SEQ ID NO: 43or SEQ ID NO: 44.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 45; SEQ ID NO:46; and SEQ ID NO: 47 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 41 or the light chainsequence of SEQ ID NO: 42.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 48; SEQ ID NO:49; and SEQ ID NO: 50 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 43 or the heavy chainsequence of SEQ ID NO: 44.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 41; the variable heavy chain region of SEQ IDNO: 43; the complementarity-determining regions (SEQ ID NO: 45; SEQ IDNO: 46; and SEQ ID NO: 47) of the variable light chain region of SEQ IDNO: 41; and the complementarity-determining regions (SEQ ID NO: 48; SEQID NO: 49; and SEQ ID NO: 50) of the variable heavy chain region of SEQID NO: 43.

In a particularly preferred embodiment of the invention, the includedchimeric anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab5, comprising, or alternatively consistingof, SEQ ID NO: 42 and SEQ ID NO: 44, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab5, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 41 and the variable heavy chain sequence of SEQID NO: 43. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 41 and/or SEQ ID NO: 43in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) of Ab5.In another embodiment of the invention, anti-NGF antibodies such as Ab5or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cells,transgenic plant or animals, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab6

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab6 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes chimeric or humanized antibodieshaving binding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 51) DIQMTQSPSTLSASVGDRVTITCQASQSIYSNLAWYQQKPGKAPKLLIYDASTLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQGFTVSDIDNA FGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 52) DIQMTQSPSTLSASVGDRVTITCQASQSIYSNLAWYQQKPGKAPKLLIYDASTLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQGFTVSDIDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 53) EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYAVGWVRQAPGKGLEWVGIIGRNGNTWYASSARGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYGRSVAYYVFNIWGPGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 54) EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYAVGWVRQAPGKGLEWVGIIGRNGNTWYASSARGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYGRSVAYYVFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ IDNO: 57 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 52, and/or oneor more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59;and SEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 54,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 51 or SEQ ID NO: 52. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 53 or SEQ ID NO: 54.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO:56; and SEQ ID NO: 57 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 52.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO:59; and SEQ ID NO: 60 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 53 or the heavy chainsequence of SEQ ID NO: 54.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 51; the variable heavy chain region of SEQ IDNO: 53; the complementarity-determining regions (SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57) of the variable light chain region of SEQ IDNO: 51; and the complementarity-determining regions (SEQ ID NO: 58; SEQID NO: 59; and SEQ ID NO: 60) of the variable heavy chain region of SEQID NO: 53.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab6, comprising, or alternatively consistingof, SEQ ID NO: 52 and SEQ ID NO: 54, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab6, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 51 and the variable heavy chain sequence of SEQID NO: 53. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 51 and/or SEQ ID NO: 53in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) of Ab6.In another embodiment of the invention, anti-NGF antibodies such as Ab6or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab7

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab7 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibits the association of NGF with p75. In oneembodiment, the invention includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 61) ADVVMTQTPASVSQPVGGTVTIKCQASEDIYNLLAWYQQKPGQPPKLLIYSASTLASGVPSRFKGSGSGTEYTLTISGLECADAATYYCQNNYLVTTYGV AFGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 62) ADVVMTQTPASVSQPVGGTVTIKCQASEDIYNLLAWYQQKPGQPPKLLIYSASTLASGVPSRFKGSGSGTEYTLTISGLECADAATYYCQNNYLVTTYGVAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 63) QEQLKESGGRLVTPGTPLTLTCTVSGFSLSSYAMIWVRQAPGKGLEYIGYIDTDTSAYYASWVKGRFTISRTSTTVDLKITSPTTEDTATYFCARSYAAY GGYPATFDPWGPGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 64) QEQLKESGGRLVTPGTPLTLTCTVSGFSLSSYAMIWVRQAPGKGLEYIGYIDTDTSAYYASWVKGRFTISRTSTTVDLKITSPTTEDTATYFCARSYAAYGGYPATFDPWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 65; SEQ ID NO: 66; and SEQ IDNO: 67 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 61 or the light chain sequence of SEQ ID NO: 62, and/or oneor more of the polypeptide sequences of SEQ ID NO: 68; SEQ ID NO: 69;and SEQ ID NO: 70 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 63 or the heavy chain sequence of SEQ ID NO: 64,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 61 or SEQ ID NO: 62. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 63 or SEQ ID NO: 64.

In a further embodiment of the invention, fragments of the antibody fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 65; SEQ ID NO: 66; andSEQ ID NO: 67 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable light chainsequence of SEQ ID NO: 61 or the light chain sequence of SEQ ID NO: 62.

In a further embodiment of the invention, fragments of the antibody fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 68; SEQ ID NO: 69; andSEQ ID NO: 70 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 63 or the heavy chain sequence of SEQ ID NO: 64.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 61; the variable heavy chain region of SEQ IDNO: 63; the complementarity-determining regions (SEQ ID NO: 65; SEQ IDNO: 66; and SEQ ID NO: 67) of the variable light chain region of SEQ IDNO: 61; and the complementarity-determining regions (SEQ ID NO: 68; SEQID NO: 69; and SEQ ID NO: 70) of the variable heavy chain region of SEQID NO: 63.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab7, comprising, or alternatively consistingof, SEQ ID NO: 62 and SEQ ID NO: 64, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab7, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 61 and the variable heavy chain sequence of SEQID NO: 63. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 61 and/or SEQ ID NO: 63in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) of Ab7.In another embodiment of the invention, anti-NGF antibodies such as Ab7or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab8

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab8 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes chimeric antibodies or humanizedhaving binding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 71) DIQMTQSPSSLSASVGDRVTITCQASEDIYNLLAWYQQKPGKVPKLLIYSASTLASGVPSRFSGSGSGTDYTLTISSLQPEDVATYYCQNNYLVTTYGVA FGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 72) DIQMTQSPSSLSASVGDRVTITCQASEDIYNLLAWYQQKPGKVPKLLIYSASTLASGVPSRFSGSGSGTDYTLTISSLQPEDVATYYCQNNYLVTTYGVAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 73) QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMIWVRQAPGKGLEYIGYIDTDTSAYYASSVKGRFTISRDNSKNTLYLQMSSLRAEDTAVYYCARSYAAYGGYPATFDPWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 74) QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMIWVRQAPGKGLEYIGYIDTDTSAYYASSVKGRFTISRDNSKNTLYLQMSSLRAEDTAVYYCARSYAAYGGYPATFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 75; SEQ ID NO: 76; and SEQ IDNO: 77 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 71 or the light chain sequence of SEQ ID NO: 72, and/or oneor more of the polypeptide sequences of SEQ ID NO: 78; SEQ ID NO: 79;and SEQ ID NO: 80 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 73 or the heavy chain sequence of SEQ ID NO: 74,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 71 or SEQ ID NO: 72. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 73 or SEQ ID NO: 74.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 75; SEQ ID NO:76; and SEQ ID NO: 77 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 71 or the light chainsequence of SEQ ID NO: 72.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 78; SEQ ID NO:79; and SEQ ID NO: 80 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 73 or the heavy chainsequence of SEQ ID NO: 74.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 71; the variable heavy chain region of SEQ IDNO: 73; the complementarity-determining regions (SEQ ID NO: 75; SEQ IDNO: 76; and SEQ ID NO: 77) of the variable light chain region of SEQ IDNO: 71; and the complementarity-determining regions (SEQ ID NO: 78; SEQID NO: 79; and SEQ ID NO: 80) of the variable heavy chain region of SEQID NO: 73.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab8, comprising, or alternatively consistingof, SEQ ID NO: 72 and SEQ ID NO: 74, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab8, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 71 and the variable heavy chain sequence of SEQID NO: 73. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 71 and/or SEQ ID NO: 73in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) of Ab8.In another embodiment of the invention, anti-NGF antibodies such as Ab8or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab9

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab9 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibits the association of NGF with p75. In oneembodiment, the invention includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 81) AYDMTQTPASVSAAVGGTVTIKCQASENIGSYLAWYQQKPGQPPELLIYRASTLASGVPSRFKGSGSGTQFTLTISGVECADAATYYCQQGYNSENLDNA FGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 82) AYDMTQTPASVSAAVGGTVTIKCQASENIGSYLAWYQQKPGQPPELLIYRASTLASGVPSRFKGSGSGTQFTLTISGVECADAATYYCQQGYNSENLDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 83) QSVEESGGRLVTPGTPLTLTCTVSGIDLSMYSMGWVRQAPGKGLEYIGWISYGGTAYYASWAKGRFTISKTSTTVELKITSPTIEDTATYFCARETPVNY YLDIWGQGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 84) QSVEESGGRLVTPGTPLTLTCTVSGIDLSMYSMGWVRQAPGKGLEYIGWISYGGTAYYASWAKGRFTISKTSTTVELKITSPTIEDTATYFCARETPVNYYLDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 85; SEQ ID NO: 86; and SEQ IDNO: 87 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 81 or the light chain sequence of SEQ ID NO: 82, and/or oneor more of the polypeptide sequences of SEQ ID NO: 88; SEQ ID NO: 89;and SEQ ID NO: 90 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 83 or the heavy chain sequence of SEQ ID NO: 84,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 81 or SEQ ID NO: 82. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 83 or SEQ ID NO: 84.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 85; SEQ ID NO:86; and SEQ ID NO: 87 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 81 or the light chainsequence of SEQ ID NO: 82.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 88; SEQ ID NO:89; and SEQ ID NO: 90 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 83 or the heavy chainsequence of SEQ ID NO: 84.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 81; the variable heavy chain region of SEQ IDNO: 83; the complementarity-determining regions (SEQ ID NO: 85; SEQ IDNO: 86; and SEQ ID NO: 87) of the variable light chain region of SEQ IDNO: 81; and the complementarity-determining regions (SEQ ID NO: 88; SEQID NO: 89; and SEQ ID NO: 90) of the variable heavy chain region of SEQID NO: 83.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab9, comprising, or alternatively consistingof, SEQ ID NO: 82 and SEQ ID NO: 84, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGor MetMab-like monovalent antibody polypeptides F. With respect toantibody Ab9, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 81 and the variable heavy chain sequence of SEQID NO: 83. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 81 and/or SEQ ID NO: 83in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) of Ab9.In another embodiment of the invention, anti-NGF antibodies such as Ab9or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab10

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab10 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 91) AYDMTQSPSSLSASVGDRVTITCQASENIGSYLAWYQQKPGKVPKLLIYRASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQGYNSENLDNA FGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 92) AYDMTQSPSSLSASVGDRVTITCQASENIGSYLAWYQQKPGKVPKLLIYRASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQGYNSENLDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 93) QVQLVESGGGVVQPGRSLRLSCAASGFTFSMYSMGWVRQAPGKGLEYIGWISYGGTAYYASSAKGRFTISRDNSKNTLYLQMSSLRAEDTAVYYCARETP VNYYLDIWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 94) QVQLVESGGGVVQPGRSLRLSCAASGFTFSMYSMGWVRQAPGKGLEYIGWISYGGTAYYASSAKGRFTISRDNSKNTLYLQMSSLRAEDTAVYYCARETPVNYYLDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 95; SEQ ID NO: 96; and SEQ IDNO: 97 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 91 or the light chain sequence of SEQ ID NO: 92, and/or oneor more of the polypeptide sequences of SEQ ID NO: 98; SEQ ID NO: 99;and SEQ ID NO: 100 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 93 or the heavy chain sequence of SEQ ID NO: 94,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 91 or SEQ ID NO: 92. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 93 or SEQ ID NO: 94.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 95; SEQ ID NO:96; and SEQ ID NO: 97 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 91 or the light chainsequence of SEQ ID NO: 92.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 98; SEQ ID NO:99; and SEQ ID NO: 100 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 93 or the heavy chainsequence of SEQ ID NO: 94.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies for treatment or preventionof pain and pain associated conditions having binding specificity to NGFcomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 91; the variable heavy chain region of SEQ IDNO: 93; the complementarity-determining regions (SEQ ID NO: 95; SEQ IDNO: 96; and SEQ ID NO: 97) of the variable light chain region of SEQ IDNO: 91; and the complementarity-determining regions (SEQ ID NO: 98; SEQID NO: 99; and SEQ ID NO: 100) of the variable heavy chain region of SEQID NO: 93.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab10, comprising, or alternatively consistingof, SEQ ID NO: 92 and SEQ ID NO: 94, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab10, the Fab fragment for treatment or prevention of pain andpain associated conditions includes the variable light chain sequence ofSEQ ID NO: 91 and the variable heavy chain sequence of SEQ ID NO: 93.This embodiment of the invention further contemplates additions,deletions, and variants of SEQ ID NO: 91 and/or SEQ ID NO: 93 in saidFab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb10. In another embodiment of the invention, anti-NGF antibodies suchas Ab10 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as bacterialor yeast cells (for example diploid yeast such as diploid Pichia) andother yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab11

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab11 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes chimeric antibodies having bindingspecificity to NGF and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 101) AFELTQTPSSVEAAVGGTVTIKCQASQNIVTNLAWYQQKPGQPPKLLIYGASTLASGVSSRFKGSGSGTQFTLTISDLECADAATYFCQSYDGFNSAGFG GGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 102) AFELTQTPSSVEAAVGGTVTIKCQASQNIVTNLAWYQQKPGQPPKLLIYGASTLASGVSSRFKGSGSGTQFTLTISDLECADAATYFCQSYDGFNSAGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 103) QSLEESGGRLVTPGTPLTLTCTASGFSLSGYDMSWVRQAPGKGLEYIGLISYDGNTYYATWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSLYAGP NAGIGPFNIWGQGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 104) QSLEESGGRLVTPGTPLTLTCTASGFSLSGYDMSWVRQAPGKGLEYIGLISYDGNTYYATWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSLYAGPNAGIGPFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 105; SEQ ID NO: 106; and SEQID NO: 107 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 101 or the light chain sequence of SEQ ID NO: 102, and/or oneor more of the polypeptide sequences of SEQ ID NO: 108; SEQ ID NO: 109;and SEQ ID NO: 110 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 103 or the heavy chain sequence of SEQ ID NO:104, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 101 or SEQ ID NO: 102. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 103 or SEQ ID NO: 104.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 105; SEQ ID NO:106; and SEQ ID NO: 107 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 101 or the light chainsequence of SEQ ID NO: 102.

In a further embodiment of the invention, fragments of the antibody fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 108; SEQ ID NO: 109; andSEQ ID NO: 110 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 103 or the heavy chain sequence of SEQ ID NO:104.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 101; the variable heavy chain region of SEQID NO: 103; the complementarity-determining regions (SEQ ID NO: 105; SEQID NO: 106; and SEQ ID NO: 107) of the variable light chain region ofSEQ ID NO: 101; and the complementarity-determining regions (SEQ ID NO:108; SEQ ID NO: 109; and SEQ ID NO: 110) of the variable heavy chainregion of SEQ ID NO: 103.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab11, comprising, or alternatively consistingof, SEQ ID NO: 102 and SEQ ID NO: 104, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab11, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 101 and the variable heavy chain sequence of SEQID NO: 103. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 101 and/or SEQ ID NO:103 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments may for treatment or prevention of pain and pain associatedconditions be produced by enzymatic digestion (e.g., papain) of Ab11. Inanother embodiment of the invention, anti-NGF antibodies such as Ab11 orFab fragments thereof may be produced via expression in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab12

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab12 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 111) AFQMTQSPSSLSASVGDRVTITCQASQNIVTNLAWYQQKPGKVPKLLIYGASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYDGFNSAGFG GGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 112) AFQMTQSPSSLSASVGDRVTITCQASQNIVTNLAWYQQKPGKVPKLLIYGASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYDGFNSAGFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 113) QVQLVESGGGVVQPGRSLRLSCAASGFSLSGYDMSWVRQAPGKGLEWVGLISYDGNTYYATSAKGRFTISRDNSKNTLYLQMSSLRAEDTAVYYCARSLYAGPNAGIGPFNIWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 114) QVQLVESGGGVVQPGRSLRLSCAASGFSLSGYDMSWVRQAPGKGLEWVGLISYDGNTYYATSAKGRFTISRDNSKNTLYLQMSSLRAEDTAVYYCARSLYAGPNAGIGPFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 115; SEQ ID NO: 116; and SEQID NO: 117 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 111 or the light chain sequence of SEQ ID NO: 112, and/or oneor more of the polypeptide sequences of SEQ ID NO: 118; SEQ ID NO: 119;and SEQ ID NO: 120 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 113 or the heavy chain sequence of SEQ ID NO:114, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 111 or SEQ ID NO: 112. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 113 or SEQ ID NO: 114.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 115; SEQ ID NO:116; and SEQ ID NO: 117 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 111 or the light chainsequence of SEQ ID NO: 112.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 118; SEQ ID NO:119; and SEQ ID NO: 120 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 113 or the heavy chainsequence of SEQ ID NO: 114.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF for treatment or prevention of pain and pain associated conditionscomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 111; the variable heavy chain region of SEQID NO: 113; the complementarity-determining regions (SEQ ID NO: 115; SEQID NO: 116; and SEQ ID NO: 117) of the variable light chain region ofSEQ ID NO: 111; and the complementarity-determining regions (SEQ ID NO:118; SEQ ID NO: 119; and SEQ ID NO: 120) of the variable heavy chainregion of SEQ ID NO: 113.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab12, comprising, or alternatively consistingof, SEQ ID NO: 112 and SEQ ID NO: 114, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab12, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 111 and the variable heavy chain sequence of SEQID NO: 113. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 111 and/or SEQ ID NO:113 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb12. In another embodiment of the invention, anti-NGF antibodies suchas Ab12 or Fab fragments thereof for treatment or prevention of pain andpain associated conditions may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab13

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude Ab13 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 121) AAVLTQTPSPVSAAVGGTVSISCQSSQNVYKNNYLSWYQQKPGQPPKLLIYKASTLASGVPSRFKGGGSGTDFTLTISDVQCDAAATYYCAGGYTSSSDN AFGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 122) AAVLTQTPSPVSAAVGGTVSISCQSSQNVYKNNYLSWYQQKPGQPPKLLIYKASTLASGVPSRFKGGGSGTDFTLTISDVQCDAAATYYCAGGYTSSSDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 123) QSVEASGGRLVTPGTPLTLTCTASGFSLSTYWMSWVRQAPGKGLEWIGDIYFSNEETNYASWAKGRFTISKTSTTVDLNVISPTTEDTATYFCARGSPDV DIGIDMWGPGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 124) QSVEASGGRLVTPGTPLTLTCTASGFSLSTYWMSWVRQAPGKGLEWIGDIYFSNEETNYASWAKGRFTISKTSTTVDLNVISPTTEDTATYFCARGSPDVDIGIDMWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 125; SEQ ID NO: 126; and SEQID NO: 127 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 121 or the light chain sequence of SEQ ID NO: 122, and/or oneor more of the polypeptide sequences of SEQ ID NO: 128; SEQ ID NO: 129;and SEQ ID NO: 130 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 123 or the heavy chain sequence of SEQ ID NO:124, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 121 or SEQ ID NO: 122. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 123 or SEQ ID NO: 124.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 125; SEQ ID NO:126; and SEQ ID NO: 127 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 121 or the light chainsequence of SEQ ID NO: 122.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 128; SEQ ID NO:129; and SEQ ID NO: 130 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 123 or the heavy chainsequence of SEQ ID NO: 124.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 121; the variable heavy chain region of SEQID NO: 123; the complementarity-determining regions (SEQ ID NO: 125; SEQID NO: 126; and SEQ ID NO: 127) of the variable light chain region ofSEQ ID NO: 121; and the complementarity-determining regions (SEQ ID NO:128; SEQ ID NO: 129; and SEQ ID NO: 130) of the variable heavy chainregion of SEQ ID NO: 123.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab13, comprising, or alternatively consistingof, SEQ ID NO: 122 and SEQ ID NO: 124, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab13, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 121 and the variable heavy chain sequence of SEQID NO: 123. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 121 and/or SEQ ID NO:123 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb13. In another embodiment of the invention, anti-NGF antibodies suchas Ab13 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as bacterialor yeast cells (for example diploid yeast such as diploid Pichia) andother yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab14

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric or humanized antibodies having binding specificity toNGF wherein the antibody is Ab14 or fragments thereof, for example asset forth below, in a therapeutically effective amount which inhibitsthe association of NGF with TrkA and further inhibits the association ofNGF with p75. In one embodiment, the invention includes chimeric orhumanized antibodies having binding specificity to NGF and possessing avariable light chain sequence comprising the sequence set forth below:

(SEQ ID NO: 131) DIQMTQSPSSLSASVGDRVTITCQSSQNVYKNNYLSWYQQKPGKVPKLLIYKASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYTSSSDN AFGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 132) DIQMTQSPSSLSASVGDRVTITCQSSQNVYKNNYLSWYQQKPGKVPKLLIYKASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYTSSSDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 133) EVQLVESGGGLVQPGGSLRLSCAASGFTVSTYWMSWVRQAPGKGLEWVGDIYFSNEETNYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGS PDVDIGIDMWGPGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 134) EVQLVESGGGLVQPGGSLRLSCAASGFTVSTYWMSWVRQAPGKGLEWVGDIYFSNEETNYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGSPDVDIGIDMWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 135; SEQ ID NO: 136; and SEQID NO: 137 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 131 or the light chain sequence of SEQ ID NO: 132, and/or oneor more of the polypeptide sequences of SEQ ID NO: 138; SEQ ID NO: 139;and SEQ ID NO: 140 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 133 or the heavy chain sequence of SEQ ID NO:134, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 131 or SEQ ID NO: 132. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 133 or SEQ ID NO: 134.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 135; SEQ ID NO:136; and SEQ ID NO: 137 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 131 or the light chainsequence of SEQ ID NO: 132.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 138; SEQ ID NO:139; and SEQ ID NO: 140 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 133 or the heavy chainsequence of SEQ ID NO: 134.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 131; the variable heavy chain region of SEQID NO: 133; the complementarity-determining regions (SEQ ID NO: 135; SEQID NO: 136; and SEQ ID NO: 137) of the variable light chain region ofSEQ ID NO: 131; and the complementarity-determining regions (SEQ ID NO:138; SEQ ID NO: 139; and SEQ ID NO: 140) of the variable heavy chainregion of SEQ ID NO: 133.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab14, comprising, or alternatively consistingof, SEQ ID NO: 132 and SEQ ID NO: 134, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab14, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 131 and the variable heavy chain sequence of SEQID NO: 133. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 131 and/or SEQ ID NO:133 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb14. In another embodiment of the invention, anti-NGF antibodies suchas Ab14 or Fab fragments thereof for treatment or prevention of pain andpain associated conditions may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab15

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is Ab15 or fragments thereof, for example as set forthbelow, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA without appreciably inhibiting theassociation of NGF with p75. In one embodiment, the invention includeschimeric antibodies for treatment or prevention of pain and painassociated conditions having binding specificity to NGF and possessing avariable light chain sequence comprising the sequence set forth below:

(SEQ ID NO: 141) AAVLTQTPSPVSAAVGDTVTIKCQSSQSVYKNNYLSWYQQKPGQPPKLLIYDASNLPSGVPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGDYDDDTDN GFGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 142) AAVLTQTPSPVSAAVGDTVTIKCQSSQSVYKNNYLSWYQQKPGQPPKLLIYDASNLPSGVPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGDYDDDTDNGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 143) QSVEESGGRLVTPGTPLTLTCTVSGIDLSSYAMIWVRQAPGKGLEYIGIIWSGGTYYATWAKGRFTISKTSTTVDLQITSPTTEDAATYFCAAGGGSIYD VWGPGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 144) QSVEESGGRLVTPGTPLTLTCTVSGIDLSSYAMIWVRQAPGKGLEYIGIIWSGGTYYATWAKGRFTISKTSTTVDLQITSPTTEDAATYFCAAGGGSIYDVWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 145; SEQ ID NO: 146; and SEQID NO: 147 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 141 or the light chain sequence of SEQ ID NO: 142, and/or oneor more of the polypeptide sequences of SEQ ID NO: 148; SEQ ID NO: 149;and SEQ ID NO: 150 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 143 or the heavy chain sequence of SEQ ID NO:144, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of,combinations of one or more of the CDRs, the variable heavy and variablelight chain sequences, and the heavy and light chain sequences set forthabove, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 141 or SEQ ID NO: 142. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 143 or SEQ ID NO: 144.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 145; SEQ ID NO:146; and SEQ ID NO: 147 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 141 or the light chainsequence of SEQ ID NO: 142.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 148; SEQ ID NO:149; and SEQ ID NO: 150 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 143 or the heavy chainsequence of SEQ ID NO: 144.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF for treatment or prevention of pain and pain associated conditionscomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 141; the variable heavy chain region of SEQID NO: 143; the complementarity-determining regions (SEQ ID NO: 145; SEQID NO: 146; and SEQ ID NO: 147) of the variable light chain region ofSEQ ID NO: 141; and the complementarity-determining regions (SEQ ID NO:148; SEQ ID NO: 149; and SEQ ID NO: 150) of the variable heavy chainregion of SEQ ID NO: 143.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab15, comprising, or alternatively consistingof, SEQ ID NO: 142 and SEQ ID NO: 144, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab15, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 141 and the variable heavy chain sequence of SEQID NO: 143. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 141 and/or SEQ ID NO:143 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb15. In another embodiment of the invention, anti-NGF antibodies fortreatment or prevention of pain and pain associated conditions such asAb15 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab16

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric or humanized antibodies having binding specificity toNGF wherein the antibody is Ab16 or fragments thereof, for example asset forth below, in a therapeutically effective amount which inhibitsthe association of NGF with TrkA without appreciably inhibiting theassociation of NGF with p75. In one embodiment, the invention includeschimeric or humanized antibodies for treatment or prevention of pain andpain associated conditions having binding specificity to NGF andpossessing a variable light chain sequence comprising the sequence setforth below:

(SEQ ID NO: 151) ALVMTQTPSSTSEPVGGTVTINCQASQNIGNDLSWYQQKPGQPPELLIYSTSKLATGVPKRFSGSRSGTQFTLTISDLECDDAATYYCLGVYSYISDDGN AFGGGTEVVVKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 152) ALVMTQTPSSTSEPVGGTVTINCQASQNIGNDLSWYQQKPGQPPELLIYSTSKLATGVPKRFSGSRSGTQFTLTISDLECDDAATYYCLGVYSYISDDGNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 153) QSVEEFGGRLVTPGTPLTLTCTVSGFSLNNYAMTWVRQAPGKGLEWIGIIGSIGTTYYASWAKGRFFISKTSTTVDLKIISPTTEDTATYFCARDAGVTV DGYGYYFNIWGPGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 154) QSVEEFGGRLVTPGTPLTLTCTVSGFSLNNYAMTWVRQAPGKGLEWIGIIGSIGTTYYASWAKGRFFISKTSTTVDLKIISPTTEDTATYFCARDAGVTVDGYGYYFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 155; SEQ ID NO: 156; and SEQID NO: 157 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 151 or the light chain sequence of SEQ ID NO: 152, and/or oneor more of the polypeptide sequences of SEQ ID NO: 158; SEQ ID NO: 159;and SEQ ID NO: 160 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 153 or the heavy chain sequence of SEQ ID NO:154, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of,combinations of one or more of the CDRs, the variable heavy and variablelight chain sequences, and the heavy and light chain sequences set forthabove, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 151 or SEQ ID NO: 152. In anotherembodiment of the invention, antibody fragments of the invention fortreatment or prevention of pain and pain associated conditions comprise,or alternatively consist of, the polypeptide sequence of SEQ ID NO: 153or SEQ ID NO: 154.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 155; SEQ ID NO:156; and SEQ ID NO: 157 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 151 or the light chainsequence of SEQ ID NO: 152.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 158; SEQ ID NO:159; and SEQ ID NO: 160 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 153 or the heavy chainsequence of SEQ ID NO: 154.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 151; the variable heavy chain region of SEQID NO: 153; the complementarity-determining regions (SEQ ID NO: 155; SEQID NO: 156; and SEQ ID NO: 157) of the variable light chain region ofSEQ ID NO: 151; and the complementarity-determining regions (SEQ ID NO:158; SEQ ID NO: 159; and SEQ ID NO: 160) of the variable heavy chainregion of SEQ ID NO: 153.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab16, comprising, or alternatively consistingof, SEQ ID NO: 152 and SEQ ID NO: 154, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab16, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 151 and the variable heavy chain sequence of SEQID NO: 153. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 151 and/or SEQ ID NO:153 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb16. In another embodiment of the invention, anti-NGF antibodies fortreatment or prevention of pain and pain associated conditions such asAb16 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab17

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is Ab17 or fragments thereof, for example as set forthbelow, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA and further inhibit the association of NGFwith p75. In one embodiment, the invention includes chimeric antibodiesfor treatment or prevention of pain and pain associated conditionshaving binding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 161) AIEMTQTPFSVSAAVGGTVTIKCQASQTISNYLAWYQQKPGQPPKLLIYGASNLESGVPSRFKGSGSGTQFTLTISDLECDDAATYYCQQGYTISNVDNN VFGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 162) AIEMTQTPFSVSAAVGGTVTIKCQASQTISNYLAWYQQKPGQPPKLLIYGASNLESGVPSRFKGSGSGTQFTLTISDLECDDAATYYCQQGYTISNVDNNVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 163) QSLEESGGRLVTPGGSLTLTCAASGFSLTGYNLVWVRQAPGKGLEWIGFISYGDTTYYASWAKGRFTISKTSTTVTLTITDLQPSDTGTYFCARETANTY DYGIWGPGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 164) QSLEESGGRLVTPGGSLTLTCAASGFSLTGYNLVWVRQAPGKGLEWIGFISYGDTTYYASWAKGRFTISKTSTTVTLTITDLQPSDTGTYFCARETANTYDYGIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 165; SEQ ID NO: 166; and SEQID NO: 167 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 161 or the light chain sequence of SEQ ID NO: 162, and/or oneor more of the polypeptide sequences of SEQ ID NO: 168; SEQ ID NO: 169;and SEQ ID NO: 170 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 163 or the heavy chain sequence of SEQ ID NO:164, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of,combinations of one or more of the CDRs, the variable heavy and variablelight chain sequences, and the heavy and light chain sequences set forthabove, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 161 or SEQ ID NO: 162. In anotheroptional embodiment of the invention, antibody fragments of theinvention comprise, or alternatively consist of, the polypeptidesequence of SEQ ID NO: 163 or SEQ ID NO: 164.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 165; SEQ ID NO:166; and SEQ ID NO: 167 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 161 or the light chainsequence of SEQ ID NO: 162.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 168; SEQ ID NO:169; and SEQ ID NO: 170 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 163 or the heavy chainsequence of SEQ ID NO: 164.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies for treatment or preventionof pain and pain associated conditions having binding specificity to NGFcomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 161; the variable heavy chain region of SEQID NO: 163; the complementarity-determining regions (SEQ ID NO: 165; SEQID NO: 166; and SEQ ID NO: 167) of the variable light chain region ofSEQ ID NO: 161; and the complementarity-determining regions (SEQ ID NO:168; SEQ ID NO: 169; and SEQ ID NO: 170) of the variable heavy chainregion of SEQ ID NO: 163.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab17, comprising, or alternatively consistingof, SEQ ID NO: 162 and SEQ ID NO: 164, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab17, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 161 and the variable heavy chain sequence of SEQID NO: 163. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 161 and/or SEQ ID NO:163 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb17. In another embodiment of the invention, anti-NGF antibodies suchas Ab17 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab18

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric or humanized antibodies having binding specificity toNGF wherein the antibody is Ab18 or fragments thereof, for example asset forth below, in a therapeutically effective amount which inhibitsthe association of NGF with TrkA and the association of NGF with p75. Inone embodiment, the invention includes chimeric or humanized antibodiesfor treatment or prevention of pain and pain associated conditionshaving binding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 171) DIQMTQSPSTLSASVGDRVTITCQASQTISNYLAWYQQKPGKAPKLLIYGASNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYTISNVDNN VFGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 172) DIQMTQSPSTLSASVGDRVTITCQASQTISNYLAWYQQKPGKAPKLLIYGASNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYTISNVDNNVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 173) EVQLVESGGGLVQPGGSLRLSCAASGFTVSGYNLVWVRQAPGKGLEWVGFISYGDTTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARETA NTYDYGIWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 174) EVQLVESGGGLVQPGGSLRLSCAASGFTVSGYNLVWVRQAPGKGLEWVGFISYGDTTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARETANTYDYGIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 175; SEQ ID NO: 176; and SEQID NO: 177 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 171 or the light chain sequence of SEQ ID NO: 172, and/or oneor more of the polypeptide sequences of SEQ ID NO: 178; SEQ ID NO: 179;and SEQ ID NO: 180 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 173 or the heavy chain sequence of SEQ ID NO:174, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of,combinations of one or more of the CDRs, the variable heavy and variablelight chain sequences, and the heavy and light chain sequences set forthabove, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 171 or SEQ ID NO: 172. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 173 or SEQ ID NO: 174.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 175; SEQ ID NO:176; and SEQ ID NO: 177 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 171 or the light chainsequence of SEQ ID NO: 172.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 178; SEQ ID NO:179; and SEQ ID NO: 180 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 173 or the heavy chainsequence of SEQ ID NO: 174.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF for treatment or prevention of pain and pain associated conditionscomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 171; the variable heavy chain region of SEQID NO: 173; the complementarity-determining regions (SEQ ID NO: 175; SEQID NO: 176; and SEQ ID NO: 177) of the variable light chain region ofSEQ ID NO: 171; and the complementarity-determining regions (SEQ ID NO:178; SEQ ID NO: 179; and SEQ ID NO: 180) of the variable heavy chainregion of SEQ ID NO: 173.

In a particularly preferred embodiment of the invention, the chimerichumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab18, comprising, or alternatively consistingof, SEQ ID NO: 172 and SEQ ID NO: 174, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for NGF orMetMab-like monovalent antibody polypeptides. With respect to antibodyAb18, the Fab fragment includes the variable light chain sequence of SEQID NO: 171 and the variable heavy chain sequence of SEQ ID NO: 173. Thisembodiment of the invention further contemplates additions, deletions,and variants of SEQ ID NO: 171 and/or SEQ ID NO: 173 in said Fab whileretaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb18. In another embodiment of the invention, anti-NGF antibodies suchas Ab18 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal or microbial systems such as bacterialor yeast cells (for example diploid yeast such as diploid Pichia) andother yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab19

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric antibodies having binding specificity to NGF whereinthe antibody is Ab19 or fragments thereof, for example as set forthbelow, in a therapeutically effective amount which inhibits theassociation of NGF with TrkA and further inhibits the association of NGFwith p75. In one embodiment, the invention includes chimeric antibodiesfor treatment or prevention of pain and pain associated conditionshaving binding specificity to NGF and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 181) AAVLTQTPSPVSAAVGGTVSISCQSSQNVYKNNYLSWYQQKPGQPPKLLIYKASTLASGVPSRFKGSGSGTDFTLTISDVQCDAAATYYCAGGYSSSSDN AFGGGTEVVVKR.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 182) AAVLTQTPSPVSAAVGGTVSISCQSSQNVYKNNYLSWYQQKPGQPPKLLIYKASTLASGVPSRFKGSGSGTDFTLTISDVQCDAAATYYCAGGYSSSSDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 183) QSVEASGGRLVMPGGSLTLTCTASGFSLSTYWMSWVRQAPGKGLEWIGDIYFSNEETNYATWAKGRFTISKTSTTVDLNVISPTTEDTATYFCARGSPDV EIAIDMWGQGTLVTVSS.

The invention also includes chimeric antibodies for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 184) QSVEASGGRLVMPGGSLTLTCTASGFSLSTYWMSWVRQAPGKGLEWIGDIYFSNEETNYATWAKGRFTISKTSTTVDLNVISPTTEDTATYFCARGSPDVEIAIDMWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 185; SEQ ID NO: 186; and SEQID NO: 187 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 181 or the light chain sequence of SEQ ID NO: 182, and/or oneor more of the polypeptide sequences of SEQ ID NO: 188; SEQ ID NO: 189;and SEQ ID NO: 190 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 183 or the heavy chain sequence of SEQ ID NO:184, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of,combinations of one or more of the CDRs, the variable heavy and variablelight chain sequences, and the heavy and light chain sequences set forthabove, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF for treatment or prevention of pain and painassociated conditions. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 181 or SEQ ID NO: 182. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 183 or SEQ ID NO: 184.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 185; SEQ ID NO:186; and SEQ ID NO: 187 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 181 or the light chainsequence of SEQ ID NO: 182.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 188; SEQ ID NO:189; and SEQ ID NO: 190 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 183 or the heavy chainsequence of SEQ ID NO: 184.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein for treatment orprevention of pain and pain associated conditions. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 181; the variable heavy chain region of SEQID NO: 183; the complementarity-determining regions (SEQ ID NO: 185; SEQID NO: 186; and SEQ ID NO: 187) of the variable light chain region ofSEQ ID NO: 181; and the complementarity-determining regions (SEQ ID NO:188; SEQ ID NO: 189; and SEQ ID NO: 190) of the variable heavy chainregion of SEQ ID NO: 183.

In a particularly preferred embodiment of the invention, the chimericanti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab19, comprising, or alternatively consistingof, SEQ ID NO: 182 and SEQ ID NO: 184, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab19, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 181 and the variable heavy chain sequence of SEQID NO: 183. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 181 and/or SEQ ID NO:183 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb19. In another embodiment of the invention, anti-NGF antibodies suchas Ab19 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab20

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric or humanized antibodies having binding specificity toNGF wherein the antibody is Ab20 or fragments thereof, for example asset forth below, in a therapeutically effective amount which inhibitsthe association of NGF with TrkA and further inhibit the association ofNGF with p75. In one embodiment, the invention includes chimeric orhumanized antibodies for treatment or prevention of pain and painassociated conditions having binding specificity to NGF and possessing avariable light chain sequence comprising the sequence set forth below:

(SEQ ID NO: 191) DIQMTQSPSSLSASVGDRVTITCQSSQNVYKNNYLSWYQQKPGKVPKLLIYKASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYTSSSDN AFGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 192) DIQMTQSPSSLSASVGDRVTITCQSSQNVYKNNYLSWYQQKPGKVPKLLIYKASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYTSSSDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 193) EVQLVESGGGLVQPGGSLRLSCAASGFTVSTYWMSWVRQAPGKGLEWVGDIYFSNEETNYATSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGS PDVEIAIDMWGQGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 194) EVQLVESGGGLVQPGGSLRLSCAASGFTVSTYWMSWVRQAPGKGLEWVGDIYFSNEETNYATSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGSPDVEIAIDMWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 195; SEQ ID NO: 196; and SEQID NO: 197 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 191 or the light chain sequence of SEQ ID NO: 192, and/or oneor more of the polypeptide sequences of SEQ ID NO: 198; SEQ ID NO: 199;and SEQ ID NO: 200 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 193 or the heavy chain sequence of SEQ ID NO:194, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of,combinations of one or more of the CDRs, the variable heavy and variablelight chain sequences, and the heavy and light chain sequences set forthabove, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 191 or SEQ ID NO: 192. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 193 or SEQ ID NO: 194.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 195; SEQ ID NO:196; and SEQ ID NO: 197 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 191 or the light chainsequence of SEQ ID NO: 192.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 198; SEQ ID NO:199; and SEQ ID NO: 200 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 193 or the heavy chainsequence of SEQ ID NO: 194.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 191; the variable heavy chain region of SEQID NO: 193; the complementarity-determining regions (SEQ ID NO: 195; SEQID NO: 196; and SEQ ID NO: 197) of the variable light chain region ofSEQ ID NO: 191; and the complementarity-determining regions (SEQ ID NO:198; SEQ ID NO: 199; and SEQ ID NO: 200) of the variable heavy chainregion of SEQ ID NO: 193.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab20, comprising, or alternatively consistingof, SEQ ID NO: 192 and SEQ ID NO: 194, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab20, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 191 and the variable heavy chain sequence of SEQID NO: 193. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 191 and/or SEQ ID NO:193 in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb20. In another embodiment of the invention, anti-NGF antibodies fortreatment or prevention of pain and pain associated conditions such asAb20 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcell, transgenic plant or animal, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab21

The invention contemplates methods of treating pain and the specificpain associated disorders alone or is association with another activeagent, e.g., an NSAID or opioid analgesic, wherein the antibodiesinclude chimeric or humanized antibodies having binding specificity toNGF wherein the antibody is Ab21 or fragments thereof, for example asset forth below, in a therapeutically effective amount which inhibitsthe association of NGF with TrkA and further inhibit the association ofNGF with p75. In one embodiment, the invention includes chimeric orhumanized antibodies having binding specificity to NGF and possessing avariable light chain sequence comprising the sequence set forth below:

(SEQ ID NO: 51) DIQMTQSPSTLSASVGDRVTITCQASQSIYSNLAWYQQKPGKAPKLLIYDASTLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQGFTVSDIDNA FGGGTKVEIKR.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 401) DIQMTQSPSTLSASVGDRVTITCQASQSIYSNLAWYQQKPGKAPKLLIYDASTLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQGFTVSDIDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a variable heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 53) EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYAVGWVRQAPGKGLEWVGIIGRNGNTWYASSARGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYGRSVAYYVFNIWGPGTLVTVSS.

The invention also includes chimeric or humanized antibodies fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF and possessing a heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 402) EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYAVGWVRQAPGKGLEWVGIIGRNGNTWYASSARGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYGRSVAYYVFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

The invention further contemplates antibodies for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ IDNO: 57 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 401, and/or oneor more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59;and SEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 402,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 51 or SEQ ID NO: 401. In anotherembodiment of the invention, antibody fragments of the invention fortreatment or prevention of pain and pain associated conditions comprise,or alternatively consist of, the polypeptide sequence of SEQ ID NO: 53or SEQ ID NO: 402.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO:56; and SEQ ID NO: 57 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 401 and monovalent antibody molecules analogousto MetMab.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to NGF for treatment or prevention of painand pain associated conditions comprise, or alternatively consist of,one or more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO:59; and SEQ ID NO: 60 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 53 or the heavy chainsequence of SEQ ID NO: 402.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 51; the variable heavy chain region of SEQ IDNO: 53; the complementarity-determining regions (SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57) of the variable light chain region of SEQ IDNO: 51; and the complementarity-determining regions (SEQ ID NO: 58; SEQID NO: 59; and SEQ ID NO: 60) of the variable heavy chain region of SEQID NO: 53.

In a particularly preferred embodiment of the invention, the chimeric orhumanized anti-NGF antibody for treatment or prevention of pain and painassociated conditions is Ab21, comprising, or alternatively consistingof, SEQ ID NO: 401 and SEQ ID NO: 402, and having at least one of thebiological activities set forth herein.

In a particularly preferred embodiment of the invention, monovalentagents are utilized in methods of treating pain in a patient withoutsubstantially increasing inflammation in said patient. Exemplarymonovalent agents include, but are not limited to, Fab, Fab′, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, IgNAR, or one or more combinations thereof.

In a further particularly preferred embodiment of the invention,antibody fragments for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, Fab(fragment antigen binding) fragments having binding specificity for NGFor MetMab-like monovalent antibody polypeptides. With respect toantibody Ab21, the Fab fragment includes the variable light chainsequence of SEQ ID NO: 51 and the variable heavy chain sequence of SEQID NO: 53. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 51 and/or SEQ ID NO: 53in said Fab while retaining binding specificity for NGF.

In one embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced by enzymatic digestion (e.g., papain) ofAb21. In another embodiment of the invention, anti-NGF antibodies fortreatment or prevention of pain and pain associated conditions such asAb21 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, plantcells, transgenic plants and animals, or microbial systems such asbacterial or yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Fragment Fab1

The invention contemplates methods of treating pain using antibodyfragment Fab1 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and further inhibit the association of NGF with p75. In oneembodiment, the invention includes Fab antibody fragments for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 405) DIQMTQSPSTLSASVGDRVTITCQASQSIYSNLAWYQQKPGKAPKLLIYDASTLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQGFTVSDIDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes Fab antibody fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 406) EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYAVGWVRQAPGKGLEWVGIIGRNGNTWYASSARGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYGRSVAYYVFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTH.

The invention further contemplates antibody fragments for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ IDNO: 57 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 405, and/or oneor more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59;and SEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 406,or combinations of these polypeptide sequences. In another embodiment ofthe invention, antibody fragments of the invention for treatment orprevention of pain and pain associated conditions comprise, oralternatively consist of, combinations of one or more of the CDRs, thevariable heavy and variable light chain sequences, and the heavy andlight chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 51 or SEQ ID NO: 405. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 53 or SEQ ID NO: 406.

In a further embodiment of the invention, antibody fragments fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; andSEQ ID NO: 57 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable light chainsequence of SEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 405.

In a further embodiment of the invention, antibody fragments fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59; andSEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 406.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 51; the variable heavy chain region of SEQ IDNO: 53; the complementarity-determining regions (SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57) of the variable light chain region of SEQ IDNO: 51; and the complementarity-determining regions (SEQ ID NO: 58; SEQID NO: 59; and SEQ ID NO: 60) of the variable heavy chain region of SEQID NO: 53.

In a particularly preferred embodiment of the invention, the anti-NGFantibody fragment for treatment or prevention of pain and painassociated conditions is Fab1, comprising SEQ ID NO: 405 and SEQ ID NO:406, and having at least one of the biological activities set forthherein. In one embodiment of the invention, antibody fragment Fab1 maybe produced by enzymatic digestion (e.g., papain) of Ab21.

Antibody Fragment Fab2

The invention contemplates methods of treating pain using antibodyfragment Fab2 or fragments thereof, for example as set forth below, in atherapeutically effective amount which inhibits the association of NGFwith TrkA and the association of NGF with p75. In one embodiment, theinvention includes Fab antibody fragments for treatment or prevention ofpain and pain associated conditions having binding specificity to NGFand possessing a light chain sequence comprising the sequence set forthbelow:

(SEQ ID NO: 407) DIQMTQSPSTLSASVGDRVTITCQASQSIYSNLAWYQQKPGKAPKLLIYDASTLESGVPSRFSGSGSGTEYTLTISSLQPDDFATYYCQQGFTVSDIDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

The invention further includes Fab antibody fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 408) EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYAVGWVRQAPGKGLEWVGIIGRNGNTWYASSARGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGYGRSVAYYVFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTH.

The invention further contemplates antibody fragments for treatment orprevention of pain and pain associated conditions comprising one or moreof the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ IDNO: 57 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 407, and/or oneor more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59;and SEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 408,or combinations of these polypeptide sequences. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, combinations of one or more of the CDRs, thevariable heavy and variable light chain sequences, and the heavy andlight chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, antibodyfragments of the invention for treatment or prevention of pain and painassociated conditions comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 51 or SEQ ID NO: 407. In anotherembodiment of the invention, antibody fragments of the invention fortreatment or prevention of pain and pain associated conditions comprise,or alternatively consist of, the polypeptide sequence of SEQ ID NO: 53or SEQ ID NO: 408.

In a further embodiment of the invention, antibody fragments fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; andSEQ ID NO: 57 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable light chainsequence of SEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 407.

In a further embodiment of the invention, antibody fragments fortreatment or prevention of pain and pain associated conditions havingbinding specificity to NGF comprise, or alternatively consist of, one ormore of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59; andSEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 408.

The invention also contemplates antibody fragments for treatment orprevention of pain and pain associated conditions which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toNGF for treatment or prevention of pain and pain associated conditionscomprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 51; the variable heavy chain region of SEQ IDNO: 53; the complementarity-determining regions (SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57) of the variable light chain region of SEQ IDNO: 51; and the complementarity-determining regions (SEQ ID NO: 58; SEQID NO: 59; and SEQ ID NO: 60) of the variable heavy chain region of SEQID NO: 53.

In a particularly preferred embodiment of the invention, the anti-NGFantibody fragment for treatment or prevention of pain and painassociated conditions is Fab2, comprising SEQ ID NO: 407 and SEQ ID NO:408, and having at least one of the biological activities set forthherein.

In another embodiment of the invention described herein (infra), Fabfragments for treatment or prevention of pain and pain associatedconditions may be produced via expression in mammalian cells such asCHO, NSO or HEK 293 cells, fungal, insect, plant cell, transgenic plantor animal, or microbial systems such as yeast cells (for example diploidyeast such as diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris. In oneembodiment of the invention, antibody fragment Fab2 may be produced byexpression in Pichia pastoris using protocols set forth herein in theexamples.

In another embodiment, antibody fragments may be present in one or moreof the following non-limiting forms: Fab, Fab′, F(ab′)₂, Fv and singlechain Fv antibody forms. In a preferred embodiment, the anti-NGFantibodies described herein further comprises the kappa constant lightchain sequence comprising the sequence set forth below:

(SEQ ID NO: 412) VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF NRGEC.

In another preferred embodiment, the anti-NGF antibodies describedherein for treatment or prevention of pain and pain associatedconditions further comprises the gamma-1 constant heavy chainpolypeptide sequence comprising the sequence set forth below:

(SEQ ID NO: 413) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention contemplates an isolated anti-NGFantibody for treatment or prevention of pain and pain associatedconditions comprising a V_(H) polypeptide sequence selected from: SEQ IDNO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153,163, 173, 183, 193, or 402, or a variant thereof; and further comprisinga V_(L) polypeptide sequence selected from: SEQ ID NO: 1, 11, 21, 31,41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181,191, or 401, or a variant thereof, wherein one or more of the frameworkresidues (FR residues) in said V_(H) or V_(L) polypeptide has beensubstituted with another amino acid residue resulting in an anti-NGFantibody that specifically binds NGF. The invention contemplateshumanized and chimeric forms of these antibodies for treatment orprevention of pain and pain associated conditions. The chimericantibodies may include an Fc derived from IgG1, IgG2, IgG3, IgG4, IgG5,IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14, IgG15, IgG16,IgG17, IgG18 or IgG19 constant regions.

In one embodiment of the invention, the antibodies or V_(H) or V_(L)polypeptides originate or are selected from one or more rabbit B cellpopulations prior to initiation of the humanization process referencedherein.

In another embodiment of the invention, the anti-NGF antibodies andfragments thereof for treatment or prevention of pain and painassociated conditions do not have binding specificity for p75 or TrkA.In a further embodiment of the invention, there is contemplated methodsfor treating pain comprising using the anti-NGF antibodies and fragmentsthereof to inhibit the association of NGF with p75 and/or TrkA. Inanother embodiment of the invention, there is contemplated methods fortreating pain comprising using anti-NGF antibodies and fragments thereofto inhibit the association of NGF with TrkA and/or multimers thereofand/or antagonizes the biological effects thereof. In another embodimentof the invention, there is contemplated methods for treating paincomprising using anti-NGF antibodies and fragments thereof to inhibitthe association of NGF with p75 and/or multimers thereof and theassociation of NGF with TrkA and/or multimers thereof, and antagonizethe biological effects of p75 and TrkA.

As stated supra, antibodies and fragments thereof may be modifiedpost-translationally to add effector moieties such as chemical linkers,detectable moieties such as for example fluorescent dyes, enzymes,substrates, bioluminescent materials, radioactive materials, andchemiluminescent moieties, or functional moieties such as for examplestreptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, andradioactive materials.

Regarding detectable moieties, further exemplary enzymes include, butare not limited to, horseradish peroxidase, acetylcholinesterase,alkaline phosphatase, beta-galactosidase and luciferase. Furtherexemplary fluorescent materials include, but are not limited to,rhodamine, fluorescein, fluorescein isothiocyanate, umbelliferone,dichlorotriazinylamine, phycoerythrin and dansyl chloride. Furtherexemplary chemiluminescent moieties include, but are not limited to,luminol. Further exemplary bioluminescent materials include, but are notlimited to, luciferin and aequorin. Further exemplary radioactivematerials include, but are not limited to, Iodine 125 (¹²⁵I), Carbon 14(¹⁴C), Sulfur 35 (³⁵S), Tritium (³H) and Phosphorus 32 (³²P).

Regarding functional moieties, exemplary cytotoxic agents include, butare not limited to, methotrexate, aminopterin, 6-mercaptopurine,6-thioguanine, cytarabine, 5-fluorouracil decarbazine; alkylating agentssuch as mechlorethamine, thioepa chlorambucil, melphalan, carmustine(BSNU), mitomycin C, lomustine (CCNU), 1-methylnitrosourea,cyclothosphamide, mechlorethamine, busulfan, dibromomannitol,streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP)cisplatin and carboplatin (paraplatin); anthracyclines includedaunorubicin (formerly daunomycin), doxorubicin (adriamycin),detorubicin, carminomycin, idarubicin, epirubicin, mitoxantrone andbisantrene; antibiotics include dactinomycin (actinomycin D), bleomycin,calicheamicin, mithramycin, and anthramycin (AMC); and antimytoticagents such as the vinca alkaloids, vincristine and vinblastine. Othercytotoxic agents include paclitaxel (taxol), ricin, pseudomonasexotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide,emetine, etoposide, tenoposide, colchicin, dihydroxy anthracin dione,1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine,propranolol, puromycin, procarbazine, hydroxyurea, asparaginase,corticosteroids, mytotane (O,P′-(DDD)), interferons, and mixtures ofthese cytotoxic agents.

Further cytotoxic agents include, but are not limited to,chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel,gemcitabine, calicheamicin, doxorubicin, 5-fluorouracil, mitomycin C,actinomycin D, cyclophosphamide, vincristine and bleomycin. Toxicenzymes from plants and bacteria such as ricin, diphtheria toxin andPseudomonas toxin may be conjugated to the humanized or chimericantibodies, or binding fragments thereof, to generatecell-type-specific-killing reagents (Youle, et al., Proc. Nat'l Acad.Sci. USA 77:5483 (1980); Gilliland, et al., Proc. Nat'l Acad. Sci. USA77:4539 (1980); Krolick, et al., Proc. Nat'l Acad. Sci. USA 77:5419(1980)).

Other cytotoxic agents include cytotoxic ribonucleases as described byGoldenberg in U.S. Pat. No. 6,653,104. Embodiments of the invention alsorelate to radioimmunoconjugates where a radionuclide that emits alpha orbeta particles is stably coupled to the antibody, or binding fragmentsthereof, with or without the use of a complex-forming agent. Suchradionuclides include beta-emitters such as Phosphorus-32 (³²P),Scandium-47 (⁴⁷Sc), Copper-67 (⁶⁷Cu), Gallium-67 (⁶⁷Ga), Yttrium-88(⁸⁸Y), Yttrium-90 (⁹⁰Y), Iodine-125 (¹²⁵I), Iodine-131 (¹³¹I),Samarium-153 (¹⁵³Sm), Lutetium-177 (¹⁷⁷Lu), Rhenium-186 (186Re) orRhenium-188 (¹⁸⁸Re), and alpha-emitters such as Astatine-211 (211At),Lead-212 (²¹²Pb), Bismuth-212 (²¹²Bi) or -213 (²¹³Bi) or Actinium-225(²²⁵Ac).

Methods are known in the art for conjugating an antibody or bindingfragment thereof to a detectable moiety and the like, such as forexample those methods described by Hunter et al, Nature 144:945 (1962);David et al, Biochemistry 13:1014 (1974); Pain et al, J. Immunol. Meth.40:219 (1981); and Nygren, J., Histochem. and Cytochem. 30:407 (1982).

Also, the antibodies or antibody fragments may be modified to affecthalf-life or circulation time such as by PEGylation. Antibodies orfragments thereof may also be chemically modified to provide additionaladvantages such as increased solubility, stability and circulating time(in vivo half-life) of the polypeptide, or decreased immunogenicity (SeeU.S. Pat. No. 4,179,337). The chemical moieties for derivatization maybe selected from water soluble polymers such as polyethylene glycol,ethylene glycol/propylene glycol copolymers, carboxymethylcellulose,dextran, polyvinyl alcohol and the like. The antibodies and fragmentsthereof may be modified at random positions within the molecule, or atpredetermined positions within the molecule and may include one, two,three or more attached chemical moieties.

The polymer may be of any molecular weight, and may be branched orunbranched. For polyethylene glycol, the preferred molecular weight isbetween about 1 kDa and about 100 kDa (the term “about” indicating thatin preparations of polyethylene glycol, some molecules will weigh more,some less, than the stated molecular weight) for ease in handling andmanufacturing. Other sizes may be used, depending on the desiredtherapeutic profile (e.g., the duration of sustained release desired,the effects, if any on biological activity, the ease in handling, thedegree or lack of antigenicity and other known effects of thepolyethylene glycol to a therapeutic protein or analog). For example,the polyethylene glycol may have an average molecular weight of about200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500,6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000,11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500,16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000,25,000, 30,000, 35,000, 40,000, 50,000, 55,000, 60,000, 65,000, 70,000,75,000, 80,000, 85,000, 90,000, 95,000, or 100,000 kDa. Branchedpolyethylene glycols are described, for example, in U.S. Pat. No.5,643,575; Morpurgo et al., Appl. Biochem. Biotechnol. 56:59-72 (1996);Vorobjev et al., Nucleosides Nucleotides 18:2745-2750 (1999); andCaliceti et al., Bioconjug. Chem. 10:638-646 (1999), the disclosures ofeach of which are incorporated herein by reference.

There are a number of attachment methods available to those skilled inthe art, See e.g., EP 0 401 384, herein incorporated by reference(coupling PEG to G-CSF), See also Malik et al., Exp. Hematol.20:1028-1035 (1992) (reporting PEGylation of GM-CSF using tresylchloride). For example, polyethylene glycol may be covalently boundthrough amino acid residues via a reactive group, such as, a free aminoor carboxyl group. Reactive groups are those to which an activatedpolyethylene glycol molecule may be bound. The amino acid residueshaving a free amino group may include lysine residues and the N-terminalamino acid residues; those having a free carboxyl group may includeaspartic acid residues glutamic acid residues and the C-terminal aminoacid residue. Sulfhydryl groups may also be used as a reactive group forattaching the polyethylene glycol molecules. Preferred for therapeuticpurposes is attachment at an amino group, such as attachment at theN-terminus or lysine group.

As suggested above, polyethylene glycol may be attached to proteins vialinkage to any of a number of amino acid residues. For example,polyethylene glycol can be linked to polypeptides via covalent bonds tolysine, histidine, aspartic acid, glutamic acid, or cysteine residues.One or more reaction chemistries may be employed to attach polyethyleneglycol to specific amino acid residues (e.g., lysine, histidine,aspartic acid, glutamic acid, or cysteine) or to more than one type ofamino acid residue (e.g., lysine, histidine, aspartic acid, glutamicacid, cysteine and combinations thereof).

Alternatively, antibodies or fragments thereof may have increased invivo half lives via fusion with albumin (including but not limited torecombinant human serum albumin or fragments or variants thereof (see,e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999, EP Patent 0 413 622,and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, herein incorporatedby reference in their entirety)) or other circulating blood proteinssuch as transferrin or ferritin. In a preferred embodiment, polypeptidesand/or antibodies of the present invention (including fragments orvariants thereof) are fused with the mature form of human serum albumin(i.e., amino acids 1-585 of human serum albumin as shown in FIGS. 1 and2 of EP Patent 0 322 094) which is herein incorporated by reference inits entirety. Polynucleotides encoding fusion proteins of the inventionare also encompassed by the invention.

Embodiments described herein further include variants and equivalentsthat are substantially homologous to the antibodies, antibody fragments,diabodies, SMIPs, camelbodies, MetMab like monovalent antibodyfragments, nanobodies, IgNAR, polypeptides, variable regions and CDRsset forth herein. These may contain, e.g., conservative substitutionmutations, (i.e., the substitution of one or more amino acids by similaramino acids). For example, conservative substitution refers to thesubstitution of an amino acid with another within the same generalclass, e.g., one acidic amino acid with another acidic amino acid, onebasic amino acid with another basic amino acid, or one neutral aminoacid by another neutral amino acid. What is intended by a conservativeamino acid substitution is well known in the art.

In another embodiment, the invention contemplates polypeptide sequenceshaving at least 90% or greater sequence homology to any one or more ofthe polypeptide sequences of antibody fragments, variable regions andCDRs set forth herein. More preferably, the invention contemplatespolypeptide sequences having at least 95, 96, 97 or 98% or greatersequence homology, even more preferably at least 98% or greater sequencehomology, and still more preferably at least 99% or greater sequencehomology to any one or more of the polypeptide sequences of antibodyfragments, variable regions and CDRs set forth herein. Methods fordetermining homology between nucleic acid and amino acid sequences arewell known to those of ordinary skill in the art.

In another embodiment, the invention further contemplates theabove-recited polypeptide homologs of the antibody fragments, variableregions and CDRs set forth herein further having anti-NGF activity.Non-limiting examples of anti-NGF activity are set forth herein.

In another embodiment, the invention further contemplates the generationand use of anti-idiotypic antibodies that bind any of the foregoingsequences. In an exemplary embodiment, such an anti-idiotypic antibodycould be administered to a subject who has received an anti-NGF antibodyto modulate, reduce, or neutralize, the effect of the anti-NGF antibody.Such anti-idiotypic antibodies could also be useful for treatment of anautoimmune disease characterized by the presence of anti-NGF antibodies.A further exemplary use of such anti-idiotypic antibodies is fordetection of the anti-NGF antibodies of the present invention, forexample to monitor the levels of the anti-NGF antibodies present in asubject's blood or other bodily fluids.

The present invention also contemplates anti-NGF antibodies comprisingany of the polypeptide or polynucleotide sequences described hereinsubstituted for any of the other polynucleotide sequences describedherein. For example, without limitation thereto, the present inventioncontemplates antibodies comprising the combination of any of thevariable light chain and variable heavy chain sequences describedherein, and further contemplates antibodies resulting from substitutionof any of the CDR sequences described herein for any of the other CDRsequences described herein.

ADDITIONAL EXEMPLARY EMBODIMENTS OF THE INVENTION

In another embodiment, the invention contemplates methods of treatingpain in an individual comprising administering to said individual one ormore anti-human NGF antibodies or antibody fragments thereof thatinhibit the association of NGF with TrkA, and/or p75, that specificallybind to the same or overlapping epitope(s) and/or compete for binding tothe same or overlapping epitope(s) on an intact human NGF polypeptide orfragment thereof as an anti-human NGF antibody selected from ananti-human NGF antibody selected from Ab1, Ab2, Ab5, Ab6, Ab7, Ab8, Ab9,Ab10, Ab11, Ab12, Ab13, Ab14, Ab17, Ab18, Ab19, Ab20, or Ab21 andfurther administering an anti-human NGF antibody or fragment thereofspecifically binds to the same or overlapping epitope(s) as an intacthuman NGF polypeptide or a fragment thereof as Ab3, Ab4, Ab15, or Ab16.

A preferred embodiment of the invention is directed to methods oftreating pain in an individual comprising administering to saidindividual chimeric or humanized antibodies and fragments thereof(including Fab fragments) having binding specificity for NGF thatinhibits the association of NGF with p75, optionally in association witha second anti-NGF antibody that further inhibits biological activitiesmediated by the binding of NGF to the TrkA receptors which substantiallydoes not affect the binding of NGF and p75. In a particularly preferredembodiment of the invention, the first chimeric or humanized anti-NGFantibodies are selected from Ab1, Ab2, Ab5-Ab14, or Ab17-Ab21 and thesecond chimeric or humanized anti-NGF antibodies are selected from Ab3,Ab4, Ab15 and Ab16.

Another preferred embodiment of the invention may further includemethods of treating or preventing pain in an individual comprisingadministering to said individual chimeric or humanized antibodies andfragments thereof (including Fab fragments) having binding specificityfor NGF that inhibit the association of NGF with TrkA that do notappreciably affect the association of NGF with p75. In this embodimentof the invention, the chimeric or humanized anti-NGF antibodies arepreferably selected from Ab3, Ab4, Ab15 and Ab16 and antibodies thatbind the same or overlapping epitope on NGF as any of these antibodies.

A preferred embodiment of the invention is directed to methods oftreating pain in an individual comprising administering to saidindividual chimeric or humanized antibodies and fragments thereofcapable of binding to NGF and selectively inhibiting biologicalactivities mediated by the binding of NGF to the TrkA receptor and thep75 receptor. In a particularly preferred embodiment of the invention,the chimeric or humanized anti-NGF antibodies which inhibit theassociation of NGF with TrkA and p75 are selected from Ab1, Ab2, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab17, Ab18, Ab19,Ab20, or Ab21. In a preferred embodiment of the invention iscontemplated a method of treating diseases or disorders associated withNGF by affecting those biological activities mediated by TrkA and/orp75. In one embodiment, the disease or disorder associated with NGF ispain. A further listing of diseases and disorders associated with NGF isprovided herein.

In another embodiment of the invention, one or more of the chimeric orhumanized anti-NGF antibodies disclosed herein, or fragments thereof(including Fab fragments) that inhibit the association of NGF with TrkAand/or p75, are utilized in methods of treating pain associated withpre- and post-operative surgery, and pain associated with trauma orinjury to the musculoskeletal system.

In another preferred embodiment of the invention, full length antibodiesand Fab fragments thereof are capable of significantly reducing pain invivo in murine models as assessed by using Gait analysis (as describedin Example 7 herein), compared to results obtained with controls, andare therefore useful in methods comprising administering said antibodiesto an individual to treat pain.

A particularly preferred embodiment of the invention contemplates theuse of Fab polypeptide sequences for the treatment of pain in a patientthat inhibit the association of NGF with TrkA and/or p75. Non-limitingtypes of pain that may be treated using Fab polypeptide sequences areprovided elsewhere in this disclosure.

In another embodiment of the invention, chimeric or humanized anti-NGFantibodies and fragments thereof (including Fab fragments) havingbinding specificity for NGF inhibit TF1 cell proliferation. In anotherembodiment of the invention, chimeric or humanized anti-NGF antibodiesand fragments thereof (including Fab fragments) having bindingspecificity for NGF inhibit PC-12 neurite outgrowth and preferablyinhibit the association of NGF with TrkA, and/or p75.

In another embodiment of the invention, the invention is directed tomethods of treating pain using anti-human NGF antibodies and/orfragments thereof which inhibit the association of NGF with TrkA, and/orp75, and which specifically binds to the same or overlapping epitopes onan intact NGF polypeptide or fragment thereof that is (are) specificallybound by Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12,Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, or Ab21 as ascertainedby epitopic mapping using overlapping linear peptide fragments whichspan the full length of the native human NGF polypeptide.

The invention is also directed to methods of treating pain using ananti-NGF antibody that binds with the same NGF epitope and/or competeswith an anti-NGF antibody for binding to the same or overlapping epitopeon NGF as an antibody or antibody fragment disclosed herein, thatinhibits the association of NGF with TrkA, and/or p75, selected fromAb1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13,Ab14, Ab17, Ab18, Ab19, Ab20 or Ab21.

In another embodiment, the invention is also directed to methods oftreating or preventing pain using an isolated anti-NGF antibody and/orantibody fragment comprising one or more of the CDRs contained in theV_(H) polypeptide sequences selected from: 3, 13, 23, 33, 43, 53, 63,73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, or 402, ora variant thereof, and/or one or more of the CDRs contained in the V_(L)polypeptide sequences selected from: 1, 11, 21, 31, 41, 51, 61, 71, 81,91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, or 401, or avariant thereof.

In one embodiment of the invention, the anti-human NGF antibodydiscussed in the two prior paragraphs comprises at least 2complementarity determining regions (CDRs) in each the variable lightand the variable heavy regions which are identical to those contained inan anti-human NGF antibody selected from the group consisting of Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14,Ab15, Ab16, Ab17, Ab18, Ab19, Ab20 or Ab21.

In a preferred embodiment, the anti-human NGF antibody discussed abovefor treatment or prevention of pain and pain associated conditionscomprises at least 2 complementarity determining regions (CDRs) in eachthe variable light and the variable heavy regions which are identical tothose contained in Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10,Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20 or Ab21. Inanother embodiment, all of the CDRs of the anti-human NGF antibodydiscussed above are identical to the CDRs contained in an anti-human NGFantibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20 orAb21. In a preferred embodiment of the invention, all of the CDRs of theanti-human NGF antibody discussed above are identical to the CDRscontained in an anti-human NGF antibody selected from Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16,Ab17, Ab18, Ab19, Ab20 or Ab21.

The invention further contemplates that the one or more anti-human NGFantibodies discussed above are aglycosylated; that contain an Fc regionthat has been modified to alter effector function, half-life,proteolysis, and/or glycosylation; are human, humanized, single chain orchimeric; and are a humanized antibody derived from a rabbit (parent)anti-human NGF antibody for treatment or prevention of pain and painassociated conditions.

The invention further contemplates one or more anti-human NGF antibodiesfor treatment or prevention of pain and pain associated conditionswherein the framework regions (FRs) in the variable light region and thevariable heavy regions of said antibody respectively are human FRs whichare unmodified or which have been modified by the substitution of atmost 2 or 3 human FR residues in the variable light or heavy chainregion with the corresponding FR residues of the parent rabbit antibody,and wherein said human FRs have been derived from human variable heavyand light chain antibody sequences which have been selected from alibrary of human germline antibody sequences based on their high levelof homology to the corresponding rabbit variable heavy or light chainregions relative to other human germline antibody sequences contained inthe library.

In one embodiment of the invention, the anti-human NGF antibody orfragment for treatment or prevention of pain and pain associatedconditions specifically binds to NGF expressing human cells and/or tocirculating soluble NGF molecules in vivo, including NGF expressed on orby human cells in a patient with a disease associated with cells thatexpress NGF and inhibits the association of NGF with TrkA and/or p75.

In another embodiment, the disease is selected from inflammatory pain,post-operative incision pain, complex regional pain syndrome, cancerpain, primary or metastatic bone cancer pain, fracture pain,osteoporotic fracture pain, pain resulting from burn, osteoporosis, goutjoint pain, pain associated with sickle cell crises, and othernociceptic pain, as well as hepatocellular carcinoma, breast cancer,liver cirrhosis, neurogenic pain, neuropathic pain, nociceptic pain,trigeminal neuralgia, post-herpetic neuralgia, phantom limb pain,fibromyalgia, menstrual pain, ovarialgia, reflex sympathetic dystrophy,neurogenic pain, osteoarthritis or rheumatoid arthritis pain, lower backpain, diabetic neuropathy, sciatica, or migraine.

The invention further contemplates anti-human NGF antibodies orfragments for treatment or prevention of pain and pain associatedconditions directly or indirectly attached to a detectable label ortherapeutic agent that preferably inhibit the association of NGF withTrkA, and/or p75.

The invention also contemplates one or more nucleic acid sequences whichresult in the expression of an anti-human NGF antibody or antibodyfragment for treatment or prevention of pain and pain associatedconditions as set forth above that preferably inhibits the associationof NGF with TrkA, and/or p75, and/or that binds NGF/TrkA complexes orNGF/p75 complexes, including those comprising, or alternativelyconsisting of, yeast or human preferred codons. The invention alsocontemplates vectors (including plasmids or recombinant viral vectors)comprising said nucleic acid sequence(s). The invention alsocontemplates host cells or recombinant host cells expressing at leastone of the antibodies set forth above, including a mammalian, yeast,bacterial, and insect cells. In a preferred embodiment, the host cell isa yeast cell. In a preferred embodiment, the yeast cell is a diploidalyeast cell. In a more preferred embodiment, the yeast cell is a Pichiayeast.

The invention also contemplates a method of treatment comprisingadministering to a patient with a disease or condition associated withNGF expressing cells a therapeutically effective amount of at least oneanti-human NGF antibody or fragment described herein that preferablyinhibits the association of NGF with TrkA, and/or p75 and/or that bindsNGF/TrkA complexes or NGF/p75 complexes. The invention also contemplatesthat the treatment method for treatment or prevention of pain and painassociated conditions may involve the administration of two or moreanti-NGF antibodies or fragments thereof and disclosed herein. If morethan one antibody is administered to the patient, the multipleantibodies may be administered simultaneously or concurrently, or may bestaggered in their administration. The diseases that may be treated arepresented in the non-limiting list set forth above and elsewhere herein.In a preferred embodiment, the disease is selected from cancer pain orneuropathic pain. In a particularly preferred embodiment, the disease iscancer pain. In another embodiment the treatment further includes theadministration of another therapeutic agent or regimen selected fromchemotherapy, radiotherapy, cytokine administration or gene therapy.

In a non-limiting embodiment of the invention, another therapeutic agentor regimen includes Taxol (paclitaxel) or its derivatives, platinumcompounds such as carboplatin or cisplatin, anthracyclines such asdoxorubicin, alkylating agents such as cyclophosphamide,anti-metabolites such as 5-fluorouracil, or etoposide.

The invention further contemplates a method of in vivo imaging whichdetects the presence of cells which express NGF comprising administeringa diagnostically effective amount of at least one anti-human NGFantibody, preferably one that inhibits the association of NGF with TrkA,and/or NGF with p75 and/or that binds NGF/TrkA complexes or NGF/p75complexes. In one embodiment, said administration further includes theadministration of a radionuclide or fluorophore that facilitatesdetection of the antibody at NGF expressing disease sites. In anotherembodiment of the invention, the method of in vivo imaging is used todetect NGF expressing tumors or metastases, or tumors or metastasesexpressing TrkA and/or p75 capable of binding to NGF or comprisingNGF/TrkA complexes or NGF/p75 complexes. In a further embodiment, theresults of said in vivo imaging method are used to facilitate the designof an appropriate therapeutic regimen, including therapeutic regimensincluding radiotherapy, chemotherapy or a combination thereof.

Polynucleotides Encoding Anti-NGF Antibody Polypeptides Antibody Ab1

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab1 polypeptides which inhibit theassociation of NGF with TrkA and further inhibit the association of NGFwith p75 for treatment or prevention of pain and pain associatedconditions having binding specificity to NGF in methods of treating painin an individual comprising administering to said individual antibodyAb1 polypeptides. In one embodiment of the invention, polynucleotides ofthe invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 1:

(SEQ ID NO: 201) GCCCTTGTGATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTCAGAACATTTACAGCAATTTAGCCTGGTATCAACAGAGACCAGGGCAGCGTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGATGCTGGGGTCCCATCGCGGTTCAGAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGACGATGTTGGCACTTACTACTGTCAAAGTGCTTTTGATAGTGATAGTACTGAAAATACTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 2:

(SEQ ID NO: 202) GCCCTTGTGATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTCAGAACATTTACAGCAATTTAGCCTGGTATCAACAGAGACCAGGGCAGCGTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGATGCTGGGGTCCCATCGCGGTTCAGAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGACGATGTTGGCACTTACTACTGTCAAAGTGCTTTTGATAGTGATAGTACTGAAAATACTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 3:

(SEQ ID NO: 203) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGCTTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTACTAGTATTGGTAGCACAGTCTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGCTACGATGACTATGATGAGATGACCTACTTTAACATCTGGGGCCAGGGGACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 4:

(SEQ ID NO: 204) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGCTTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTACTAGTATTGGTAGCACAGTCTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGCTACGATGACTATGATGAGATGACCTACTTTAACATCTGGGGCCAGGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 205; SEQ ID NO: 206; and SEQ ID NO: 207 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 1 or the light chain sequence of SEQ ID NO: 2.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 208; SEQ ID NO: 209; and SEQ ID NO: 210 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 3 or the heavy chain sequence of SEQ ID NO: 4.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 201 encoding the light chain variable sequenceof SEQ ID NO: 1; the polynucleotide SEQ ID NO: 202 encoding the lightchain sequence of SEQ ID NO: 2; the polynucleotide SEQ ID NO: 203encoding the heavy chain variable sequence of SEQ ID NO: 3; thepolynucleotide SEQ ID NO: 204 encoding the heavy chain sequence of SEQID NO: 4; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 205; SEQ ID NO: 206; and SEQ ID NO: 207) of thelight chain variable sequence of SEQ ID NO: 1 or the light chainsequence of SEQ ID NO: 2; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 208; SEQ ID NO: 209; andSEQ ID NO: 210) of the heavy chain variable sequence of SEQ ID NO: 3 orthe heavy chain sequence of SEQ ID NO: 4.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity for NGF. With respect to antibody Ab1, the polynucleotidesencoding the full length Ab1 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 202 encoding the light chain sequenceof SEQ ID NO: 2 and the polynucleotide SEQ ID NO: 204 encoding the heavychain sequence of SEQ ID NO: 4.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments for treatment orprevention of pain and pain associated conditions may be produced byenzymatic digestion (e.g., papain) of Ab1 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab1 or Fab fragments thereofmay be produced via expression of Ab1 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab2

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab2 polypeptides which inhibit theassociation of NGF with TrkA and further inhibit the association of NGFwith p75 for treatment or prevention of pain and pain associatedconditions having binding specificity to NGF in methods of treating painin an individual comprising administering to said individual antibodyAb2 polypeptides. The invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to NGF fortreatment or prevention of pain and pain associated conditions. In oneembodiment of the invention, polynucleotides of the invention comprise,or alternatively consist of, the following polynucleotide sequenceencoding the variable light chain polypeptide sequence of SEQ ID NO: 11:

(SEQ ID NO: 211) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAACATTTACAGCAACTTAGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGGTGCATCCAATCTGGATGCTGGAGTCCCATCAAGGTTCTCTGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAAAGTGCTTTTGATAGTGATAGTACTGAAAACACTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 12:

(SEQ ID NO: 212) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAACATTTACAGCAACTTAGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGGTGCATCCAATCTGGATGCTGGAGTCCCATCAAGGTTCTCTGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAAAGTGCTTTTGATAGTGATAGTACTGAAAACACTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 13:

(SEQ ID NO: 213) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAGCTATGCAATGAGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTACTAGTATTGGTAGCACAGTCTACGCGAGCAGCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGCTACGATGACTATGATGAGATGACCTACTTTAACATCTGGGGCCAAGGGACCCTCGT CACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 14:

(SEQ ID NO: 214) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAGCTATGCAATGAGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTACTAGTATTGGTAGCACAGTCTACGCGAGCAGCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGCTACGATGACTATGATGAGATGACCTACTTTAACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 215; SEQ ID NO: 216; and SEQ ID NO: 217 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 11 or the light chain sequence of SEQ ID NO: 12.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 218; SEQ ID NO: 219; and SEQ ID NO: 220 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 13 or the heavy chain sequence of SEQ ID NO: 14.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 211 encoding the light chain variable sequenceof SEQ ID NO: 11; the polynucleotide SEQ ID NO: 212 encoding the lightchain sequence of SEQ ID NO: 12; the polynucleotide SEQ ID NO: 213encoding the heavy chain variable sequence of SEQ ID NO: 13; thepolynucleotide SEQ ID NO: 214 encoding the heavy chain sequence of SEQID NO: 14; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 215; SEQ ID NO: 216; and SEQ ID NO: 217) of thelight chain variable sequence of SEQ ID NO: 11 or the light chainsequence of SEQ ID NO: 12; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 218; SEQ ID NO: 219; andSEQ ID NO: 220) of the heavy chain variable sequence of SEQ ID NO: 13 orthe heavy chain sequence of SEQ ID NO: 14.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity for NGF. With respect to antibody Ab2, the polynucleotidesencoding the full length Ab2 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 212 encoding the light chain sequenceof SEQ ID NO: 12 and the polynucleotide SEQ ID NO: 214 encoding theheavy chain sequence of SEQ ID NO: 14.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab2 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab2 or Fab fragments thereofmay be produced via expression of Ab2 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab3

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab3 polypeptides which inhibit theassociation of NGF with TrkA and do not appreciably inhibit theassociation of NGF with p75, having binding specificity to NGF inmethods of treating pain in an individual comprising administering tosaid individual antibody Ab3 polypeptides. The invention is furtherdirected to polynucleotides encoding antibody polypeptides havingbinding specificity to NGF. In one embodiment of the invention,polynucleotides of the invention comprise, or alternatively consist of,the following polynucleotide sequence encoding the variable light chainpolypeptide sequence of SEQ ID NO: 21:

(SEQ ID NO: 221) GCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCAGCTATGGGAGACACAGTCACCATCAAGTGCCAGTCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAGGCTCCTGATCTATGATGCATCCAATCTGCCATCTGGGGTCCCATCACGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGATTATGATGATGATGCTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 22:

(SEQ ID NO: 222) GCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCAGCTATGGGAGACACAGTCACCATCAAGTGCCAGTCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAGGCTCCTGATCTATGATGCATCCAATCTGCCATCTGGGGTCCCATCACGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGATTATGATGATGATGCTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 23:

(SEQ ID NO: 223) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTAGCTATGTAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGAATCACTTGGAGTGCTGGTACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCGGAGGTGGTGGTAGTATTTATGATATTTGGGGCCCGGGCACCCTGGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 24:

(SEQ ID NO: 224) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTAGCTATGTAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGAATCACTTGGAGTGCTGGTACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCGGAGGTGGTGGTAGTATTTATGATATTTGGGGCCCGGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 225; SEQ ID NO: 226; and SEQ ID NO: 227 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 21 or the light chain sequence of SEQ ID NO: 22.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 228; SEQ ID NO: 229; and SEQ ID NO: 230 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 23 or the heavy chain sequence of SEQ ID NO: 24.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein for treatment or prevention of pain and pain associatedconditions. In one embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 221 encoding the light chain variable sequenceof SEQ ID NO: 21; the polynucleotide SEQ ID NO: 222 encoding the lightchain sequence of SEQ ID NO: 22; the polynucleotide SEQ ID NO: 223encoding the heavy chain variable sequence of SEQ ID NO: 23; thepolynucleotide SEQ ID NO: 224 encoding the heavy chain sequence of SEQID NO: 24; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 225; SEQ ID NO: 226; and SEQ ID NO: 227) of thelight chain variable sequence of SEQ ID NO: 21 or the light chainsequence of SEQ ID NO: 22; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 228; SEQ ID NO: 229; andSEQ ID NO: 230) of the heavy chain variable sequence of SEQ ID NO: 23 orthe heavy chain sequence of SEQ ID NO: 24.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity for NGF. With respect to antibody Ab3, the polynucleotidesencoding the full length Ab3 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 222 encoding the light chain sequenceof SEQ ID NO: 22 and the polynucleotide SEQ ID NO: 224 encoding theheavy chain sequence of SEQ ID NO: 24.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab3 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab3 or Fab fragments thereofmay be produced via expression of Ab3 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab4

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab4 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA and donot appreciably inhibit the association of NGF with p75, in methods oftreating pain in an individual comprising administering to saidindividual antibody Ab4 polypeptides. The invention is further directedto polynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 31:

(SEQ ID NO: 231) .GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTCTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCAATCTGCCATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGAACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCTAGGCGATTATGATGATGATGCTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 32:

(SEQ ID NO: 232) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTCTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCAATCTGCCATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGAACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCTAGGCGATTATGATGATGATGCTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 33:

(SEQ ID NO: 233) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAGCTATGTAATGATCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAATCACTTGGAGTGCTGGTACATACTACGCGAGCAGTGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTGGAGGTGGTGGTAGTATCTATGATATTTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 34:

(SEQ ID NO: 234) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAGCTATGTAATGATCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAATCACTTGGAGTGCTGGTACATACTACGCGAGCAGTGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTGGAGGTGGTGGTAGTATCTATGATATTTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 235; SEQ ID NO: 236; and SEQ ID NO: 237 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 31 or the light chain sequence of SEQ ID NO: 32.

In a further embodiment of the invention, polynucleotides encodingantibody fragments h for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 238; SEQ ID NO: 239; and SEQ ID NO: 240 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 33 or the heavy chain sequence of SEQ ID NO: 34.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 231 encoding the light chain variable sequenceof SEQ ID NO: 31; the polynucleotide SEQ ID NO: 232 encoding the lightchain sequence of SEQ ID NO: 32; the polynucleotide SEQ ID NO: 233encoding the heavy chain variable sequence of SEQ ID NO: 33; thepolynucleotide SEQ ID NO: 234 encoding the heavy chain sequence of SEQID NO: 34; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 235; SEQ ID NO: 236; and SEQ ID NO: 237) of thelight chain variable sequence of SEQ ID NO: 31 or the light chainsequence of SEQ ID NO: 32; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 238; SEQ ID NO: 239; andSEQ ID NO: 240) of the heavy chain variable sequence of SEQ ID NO: 33 orthe heavy chain sequence of SEQ ID NO: 34.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity for NGF. With respect to antibody Ab4, the polynucleotidesencoding the full length Ab4 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 232 encoding the light chain sequenceof SEQ ID NO: 32 and the polynucleotide SEQ ID NO: 234 encoding theheavy chain sequence of SEQ ID NO: 34.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab4 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies for treatment or prevention of painand pain associated conditions such as Ab4 or Fab fragments thereof maybe produced via expression of Ab4 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab5

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab5 polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75 in methods of treatingpain in an individual comprising administering to said individualantibody Ab5 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 41:

(SEQ ID NO: 241) GCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAGCATTTACAGCAATTTAGCCTGGTATCAGCAGAGACCAGGGCAGCCTCCCAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGGCGTGGAGTGTGCCGATGCTGCCTCTTACTACTGTCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 42:

(SEQ ID NO: 242) GCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAGCATTTACAGCAATTTAGCCTGGTATCAGCAGAGACCAGGGCAGCCTCCCAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGGCGTGGAGTGTGCCGATGCTGCCTCTTACTACTGTCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 43:

(SEQ ID NO: 243) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTAACTATGCAGTGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTGGGCAAGAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAAGCGAGGACACGGCCACATATTTCTGTGCCAGAGGATATGGCCGTAGTGTTGCTTATTACGTCTTTAACATCTGGGGCCCAGGCACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 44:

(SEQ ID NO: 244) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTAACTATGCAGTGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTGGGCAAGAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAAGCGAGGACACGGCCACATATTTCTGTGCCAGAGGATATGGCCGTAGTGTTGCTTATTACGTCTTTAACATCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encoding fortreatment or prevention of pain and pain associated conditions fragmentshaving binding specificity to NGF comprise, or alternatively consist of,one or more of the polynucleotide sequences of SEQ ID NO: 245; SEQ IDNO: 246; and SEQ ID NO: 247 which correspond to polynucleotides encodingthe complementarity-determining regions (CDRs, or hypervariable regions)of the light chain variable sequence of SEQ ID NO: 41 or the light chainsequence of SEQ ID NO: 42.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 248; SEQ ID NO: 249; and SEQ ID NO: 250 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 43 or the heavy chain sequence of SEQ ID NO: 44.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 241 encoding the light chain variable sequenceof SEQ ID NO: 41; the polynucleotide SEQ ID NO: 242 encoding the lightchain sequence of SEQ ID NO: 42; the polynucleotide SEQ ID NO: 243encoding the heavy chain variable sequence of SEQ ID NO: 43; thepolynucleotide SEQ ID NO: 244 encoding the heavy chain sequence of SEQID NO: 44; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 245; SEQ ID NO: 246; and SEQ ID NO: 247) of thelight chain variable sequence of SEQ ID NO: 41 or the light chainsequence of SEQ ID NO: 42; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 248; SEQ ID NO: 249; andSEQ ID NO: 250) of the heavy chain variable sequence of SEQ ID NO: 43 orthe heavy chain sequence of SEQ ID NO: 44.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab5, the polynucleotidesencoding the full length Ab5 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 242 encoding the light chain sequenceof SEQ ID NO: 42 and the polynucleotide SEQ ID NO: 244 encoding theheavy chain sequence of SEQ ID NO: 44.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab5 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab5 or Fab fragments thereofmay be produced via expression of Ab5 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab6

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab6 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab6 polypeptides. The invention is further directed topolynucleotides encoding for treatment or prevention of pain and painassociated conditions polypeptides having binding specificity to NGF. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 51:

(SEQ ID NO: 251) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTTACAGCAATCTTGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 52:

(SEQ ID NO: 252) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTTACAGCAATCTTGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 53:

(SEQ ID NO: 253) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAACTATGCAGTGGGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTCTGCAAGAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGATATGGCCGTAGTGTTGCTTATTACGTCTTTAACATCTGGGGCCCAGGGACCCTCGT CACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 54:

(SEQ ID NO: 254) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAACTATGCAGTGGGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTCTGCAAGAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGATATGGCCGTAGTGTTGCTTATTACGTCTTTAACATCTGGGGCCCAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 51 or the light chain sequence of SEQ ID NO: 52.

In a further embodiment of the invention, polynucleotides encoding fortreatment or prevention of pain and pain associated conditions fragmentshaving binding specificity to NGF comprise, or alternatively consist of,one or more of the polynucleotide sequences of SEQ ID NO: 258; SEQ IDNO: 259; and SEQ ID NO: 260 which correspond to polynucleotides encodingthe complementarity-determining regions (CDRs, or hypervariable regions)of the heavy chain variable sequence of SEQ ID NO: 53 or the heavy chainsequence of SEQ ID NO: 54.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 251 encoding the light chain variable sequenceof SEQ ID NO: 51; the polynucleotide SEQ ID NO: 252 encoding the lightchain sequence of SEQ ID NO: 52; the polynucleotide SEQ ID NO: 253encoding the heavy chain variable sequence of SEQ ID NO: 53; thepolynucleotide SEQ ID NO: 254 encoding the heavy chain sequence of SEQID NO: 54; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257) of thelight chain variable sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 52; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 258; SEQ ID NO: 259; andSEQ ID NO: 260) of the heavy chain variable sequence of SEQ ID NO: 53 orthe heavy chain sequence of SEQ ID NO: 54.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab6, the polynucleotidesencoding the full length Ab6 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 252 encoding the light chain sequenceof SEQ ID NO: 52 and the polynucleotide SEQ ID NO: 254 encoding theheavy chain sequence of SEQ ID NO: 54.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab6 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab6 or Fab fragments thereofmay be produced via expression of Ab6 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab7

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab7 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab7 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 61:

(SEQ ID NO: 261) GCCGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTCAACCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGGACATTTATAACTTATTGGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGGCCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAAAACAATTATCTTGTTACTACTTATGGTGTTGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 62:

(SEQ ID NO: 262) GCCGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTCAACCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGGACATTTATAACTTATTGGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGGCCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAAAACAATTATCTTGTTACTACTTATGGTGTTGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 63:

(SEQ ID NO: 263) CAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGTACAGTCTCTGGATTCTCCCTCAGTAGCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATACATTGATACTGATACTAGCGCATACTACGCGAGCTGGGTGAAAGGCCGATTCACCATCTCCAGAACCTCGACCACGGTGGATCTCAAAATCACTAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGATCTTATGCTGCTTATGGTGGTTATCCTGCTACTTTTGATCCCTGGGGCCCAGGCACCCTGGTCAC CGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 64:

(SEQ ID NO: 264) CAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGTACAGTCTCTGGATTCTCCCTCAGTAGCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATACATTGATACTGATACTAGCGCATACTACGCGAGCTGGGTGAAAGGCCGATTCACCATCTCCAGAACCTCGACCACGGTGGATCTCAAAATCACTAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGATCTTATGCTGCTTATGGTGGTTATCCTGCTACTTTTGATCCCTGGGGCCCAGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 265; SEQ ID NO: 266; and SEQ ID NO: 267 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 61 or the light chain sequence of SEQ ID NO: 62.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 268; SEQ ID NO: 269; and SEQ ID NO: 270 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 63 or the heavy chain sequence of SEQ ID NO: 64.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 261 encoding the light chain variable sequenceof SEQ ID NO: 61; the polynucleotide SEQ ID NO: 262 encoding the lightchain sequence of SEQ ID NO: 62; the polynucleotide SEQ ID NO: 263encoding the heavy chain variable sequence of SEQ ID NO: 63; thepolynucleotide SEQ ID NO: 264 encoding the heavy chain sequence of SEQID NO: 64; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 265; SEQ ID NO: 266; and SEQ ID NO: 267) of thelight chain variable sequence of SEQ ID NO: 61 or the light chainsequence of SEQ ID NO: 62; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 268; SEQ ID NO: 269; andSEQ ID NO: 270) of the heavy chain variable sequence of SEQ ID NO: 63 orthe heavy chain sequence of SEQ ID NO: 64.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab7, the polynucleotidesencoding the full length Ab7 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 262 encoding the light chain sequenceof SEQ ID NO: 62 and the polynucleotide SEQ ID NO: 264 encoding theheavy chain sequence of SEQ ID NO: 64.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab7 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab7 or Fab fragments thereofmay be produced via expression of Ab7 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab8

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab8 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit he association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab8 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 71:

(SEQ ID NO: 271) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTGAGGACATTTACAACTTATTGGCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTACACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAACAACTATCTTGTTACTACTTATGGTGTTGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 72:

(SEQ ID NO: 272) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTGAGGACATTTACAACTTATTGGCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTACACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAACAACTATCTTGTTACTACTTATGGTGTTGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 73:

(SEQ ID NO: 273) CAGGTACAGCTGGTGGAGTCTGGTGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCTTCTGGATTCACCTTCAGTAGCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATACATTGATACTGATACTAGCGCATACTACGCAAGCAGTGTGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTACCTGCAAATGTCTAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCTAGATCTTATGCTGCTTATGGTGGTTATCCTGCTACTTTTGATCCCTGGGGCCAAGGTACCCT CGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 74:

(SEQ ID NO: 274) CAGGTACAGCTGGTGGAGTCTGGTGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCTTCTGGATTCACCTTCAGTAGCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATACATTGATACTGATACTAGCGCATACTACGCAAGCAGTGTGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTACCTGCAAATGTCTAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCTAGATCTTATGCTGCTTATGGTGGTTATCCTGCTACTTTTGATCCCTGGGGCCAAGGTACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCG GGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 275; SEQ ID NO: 276; and SEQ ID NO: 277 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 71 or the light chain sequence of SEQ ID NO: 72.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 278; SEQ ID NO: 279; and SEQ ID NO: 280 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 73 or the heavy chain sequence of SEQ ID NO: 74.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 271 encoding the light chain variable sequenceof SEQ ID NO: 71; the polynucleotide SEQ ID NO: 272 encoding the lightchain sequence of SEQ ID NO: 72; the polynucleotide SEQ ID NO: 273encoding the heavy chain variable sequence of SEQ ID NO: 73; thepolynucleotide SEQ ID NO: 274 encoding the heavy chain sequence of SEQID NO: 74; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 275; SEQ ID NO: 276; and SEQ ID NO: 277) of thelight chain variable sequence of SEQ ID NO: 71 or the light chainsequence of SEQ ID NO: 72; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 278; SEQ ID NO: 279; andSEQ ID NO: 280) of the heavy chain variable sequence of SEQ ID NO: 73 orthe heavy chain sequence of SEQ ID NO: 74.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab8, the polynucleotidesencoding the full length Ab8 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 272 encoding the light chain sequenceof SEQ ID NO: 72 and the polynucleotide SEQ ID NO: 274 encoding theheavy chain sequence of SEQ ID NO: 74.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as bacterial or yeast cellssuch as the yeast Pichia. Suitable Pichia species include, but are notlimited to, Pichia pastoris. In one embodiment of the inventiondescribed herein (infra), Fab fragments may be produced by enzymaticdigestion (e.g., papain) of Ab8 following expression of the full-lengthpolynucleotides in a suitable host. In another embodiment of theinvention, anti-NGF antibodies such as Ab8 or Fab fragments thereof maybe produced via expression of Ab8 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab9

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab9 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab9 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 81:

(SEQ ID NO: 281) GCCTATGATATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGAACATTGGTAGCTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCGAACTCCTGATCTACAGGGCGTCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAACAGGGTTATAATAGTGAGAATCTTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 82:

(SEQ ID NO: 282) GCCTATGATATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGAACATTGGTAGCTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCGAACTCCTGATCTACAGGGCGTCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAACAGGGTTATAATAGTGAGAATCTTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 83:

(SEQ ID NO: 283) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGACCTCAGTATGTATTCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATGGATTAGTTATGGTGGTACTGCATATTACGCGAGCTGGGCGAAGGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGAGCTGAAGATCACCAGTCCGACAATCGAGGACACGGCCACCTATTTCTGTGCCAGAGAGACTCCTGTTAATTATTATTTGGACATTTGGGGCCAGGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 84:

(SEQ ID NO: 284) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGACCTCAGTATGTATTCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATGGATTAGTTATGGTGGTACTGCATATTACGCGAGCTGGGCGAAGGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGAGCTGAAGATCACCAGTCCGACAATCGAGGACACGGCCACCTATTTCTGTGCCAGAGAGACTCCTGTTAATTATTATTTGGACATTTGGGGCCAGGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCAcCCTCCTCCaAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 285; SEQ ID NO: 286; and SEQ ID NO: 287 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 81 or the light chain sequence of SEQ ID NO: 82.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 288; SEQ ID NO: 289; and SEQ ID NO: 290 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 83 or the heavy chain sequence of SEQ ID NO: 84.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 281 encoding the light chain variable sequenceof SEQ ID NO: 81; the polynucleotide SEQ ID NO: 282 encoding the lightchain sequence of SEQ ID NO: 82; the polynucleotide SEQ ID NO: 283encoding the heavy chain variable sequence of SEQ ID NO: 83; thepolynucleotide SEQ ID NO: 284 encoding the heavy chain sequence of SEQID NO: 84; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 285; SEQ ID NO: 286; and SEQ ID NO: 287) of thelight chain variable sequence of SEQ ID NO: 81 or the light chainsequence of SEQ ID NO: 82; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 288; SEQ ID NO: 289; andSEQ ID NO: 290) of the heavy chain variable sequence of SEQ ID NO: 83 orthe heavy chain sequence of SEQ ID NO: 84.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab9, the polynucleotidesencoding the full length Ab9 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 282 encoding the light chain sequenceof SEQ ID NO: 82 and the polynucleotide SEQ ID NO: 284 encoding theheavy chain sequence of SEQ ID NO: 84.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab9 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab9 or Fab fragments thereofmay be produced via expression of Ab9 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab10

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab10 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab10 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 91:

(SEQ ID NO: 291) GCCTATGATATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTGAGAACATTGGTAGCTACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATAGGGCTTCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAACAGGGTTACAATAGTGAGAATCTTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 92:

(SEQ ID NO: 292) GCCTATGATATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTGAGAACATTGGTAGCTACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATAGGGCTTCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAACAGGGTTACAATAGTGAGAATCTTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 93:

(SEQ ID NO: 293) CAGGTACAGCTGGTGGAGTCTGGTGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCTTCTGGATTCACCTTCAGTATGTATTCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATGGATTAGTTATGGTGGTACTGCATACTACGCTAGCAGCGCTAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTACCTGCAAATGTCTAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCTAGAGAGACTCCTGTTAATTACTACTTGGACATTTGGGGCCAAGGTACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 94:

(SEQ ID NO: 294) CAGGTACAGCTGGTGGAGTCTGGTGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCTTCTGGATTCACCTTCAGTATGTATTCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATGGATTAGTTATGGTGGTACTGCATACTACGCTAGCAGCGCTAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTACCTGCAAATGTCTAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCTAGAGAGACTCCTGTTAATTACTACTTGGACATTTGGGGCCAAGGTACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 295; SEQ ID NO: 296; and SEQ ID NO: 297 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 91 or the light chain sequence of SEQ ID NO: 92.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 298; SEQ ID NO: 299; and SEQ ID NO: 300 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 93 or the heavy chain sequence of SEQ ID NO: 94.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 291 encoding the light chain variable sequenceof SEQ ID NO: 91; the polynucleotide SEQ ID NO: 292 encoding the lightchain sequence of SEQ ID NO: 92; the polynucleotide SEQ ID NO: 293encoding the heavy chain variable sequence of SEQ ID NO: 93; thepolynucleotide SEQ ID NO: 294 encoding the heavy chain sequence of SEQID NO: 94; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 295; SEQ ID NO: 296; and SEQ ID NO: 297) of thelight chain variable sequence of SEQ ID NO: 91 or the light chainsequence of SEQ ID NO: 92; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 298; SEQ ID NO: 299; andSEQ ID NO: 300) of the heavy chain variable sequence of SEQ ID NO: 93 orthe heavy chain sequence of SEQ ID NO: 94.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab10, the polynucleotidesencoding the full length Ab10 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 292 encoding the light chainsequence of SEQ ID NO: 92 and the polynucleotide SEQ ID NO: 294 encodingthe heavy chain sequence of SEQ ID NO: 94.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as bacterial or yeast cellssuch as the yeast Pichia. Suitable Pichia species include, but are notlimited to, Pichia pastoris. In one embodiment of the inventiondescribed herein (infra), Fab fragments may be produced by enzymaticdigestion (e.g., papain) of Ab10 following expression of the full-lengthpolynucleotides in a suitable host. In another embodiment of theinvention, anti-NGF antibodies such as Ab10 or Fab fragments thereof maybe produced via expression of Ab10 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab11

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab11 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab11 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 101:

(SEQ ID NO: 301) GCATTCGAATTGACCCAGACTCCATCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAACATTGTTACCAATTTAGCCTGGTATCAACAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTATTTCTGTCAGAGCTATGATGGTTTTAATAGTGCTGGGTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:102:

(SEQ ID NO: 302) GCATTCGAATTGACCCAGACTCCATCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAACATTGTTACCAATTTAGCCTGGTATCAACAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTATTTCTGTCAGAGCTATGATGGTTTTAATAGTGCTGGGTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTA G.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 103:

(SEQ ID NO: 303) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTGGCTACGACATGAGCTGGGTCCGCCAGGCTCCAGGAAAGGGGCTGGAATACATCGGACTCATTAGTTATGATGGTAACACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAAGTCTTTATGCTGGTCCTAATGCTGGTATCGGACCGTTTAACATCTGGGGCCAGGGGACCCTCGT CACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:104:

(SEQ ID NO: 304) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTGGCTACGACATGAGCTGGGTCCGCCAGGCTCCAGGAAAGGGGCTGGAATACATCGGACTCATTAGTTATGATGGTAACACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAAGTCTTTATGCTGGTCCTAATGCTGGTATCGGACCGTTTAACATCTGGGGCCAGGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCAcCCTCCTCCaAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGaTCTCCCgGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 305; SEQ ID NO: 306; and SEQ ID NO: 307 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 101 or the light chain sequence of SEQ ID NO: 102.

In a further embodiment of the invention, polynucleotides encoding fortreatment or prevention of pain and pain associated conditions fragmentshaving binding specificity to NGF comprise, or alternatively consist of,one or more of the polynucleotide sequences of SEQ ID NO: 308; SEQ IDNO: 309; and SEQ ID NO: 310 which correspond to polynucleotides encodingthe complementarity-determining regions (CDRs, or hypervariable regions)of the heavy chain variable sequence of SEQ ID NO: 103 or the heavychain sequence of SEQ ID NO: 104.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 301 encoding the light chain variable sequenceof SEQ ID NO: 101; the polynucleotide SEQ ID NO: 302 encoding the lightchain sequence of SEQ ID NO: 102; the polynucleotide SEQ ID NO: 303encoding the heavy chain variable sequence of SEQ ID NO: 103; thepolynucleotide SEQ ID NO: 304 encoding the heavy chain sequence of SEQID NO: 104; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 305; SEQ ID NO: 306; and SEQ ID NO: 307) of thelight chain variable sequence of SEQ ID NO: 101 or the light chainsequence of SEQ ID NO: 102; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 308; SEQ ID NO: 309; andSEQ ID NO: 310) of the heavy chain variable sequence of SEQ ID NO: 103or the heavy chain sequence of SEQ ID NO: 104.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab11, the polynucleotidesencoding the full length Ab11 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 302 encoding the light chainsequence of SEQ ID NO: 102 and the polynucleotide SEQ ID NO: 304encoding the heavy chain sequence of SEQ ID NO: 104.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab11 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab11 or Fab fragments thereofmay be produced via expression of Ab11 polynucleotides in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab12

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab12 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andthe association of NGF with p75, in methods of treating pain in anindividual comprising administering to said individual antibody Ab12polypeptides. The invention is further directed to polynucleotidesencoding antibody polypeptides for treatment or prevention of pain andpain associated conditions having binding specificity to NGF. In oneembodiment of the invention, polynucleotides of the invention comprise,or alternatively consist of, the following polynucleotide sequenceencoding the variable light chain polypeptide sequence of SEQ ID NO:111:

(SEQ ID NO: 311) GCATTCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAACATTGTTACCAACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAGAGCTATGATGGTTTCAATAGTGCTGGTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:112:

(SEQ ID NO: 312) GCATTCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAACATTGTTACCAACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAGAGCTATGATGGTTTCAATAGTGCTGGTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTA G.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 113:

(SEQ ID NO: 313) CAGGTACAGCTGGTGGAGTCTGGTGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCTTCTGGATTCTCCCTCAGTGGCTACGACATGAGCTGGGTCCGTCAGGCTCCAGGCAAGGGACTGGAGTGGGTGGGACTCATTAGTTATGATGGTAACACATACTACGCGACCTCCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTACCTGCAAATGTCTAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCTAGAAGTCTTTATGCTGGTCCTAATGCTGGTATCGGACCGTTTAACATCTGGGGCCAAGGTACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:114:

(SEQ ID NO: 314) CAGGTACAGCTGGTGGAGTCTGGTGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCTTCTGGATTCTCCCTCAGTGGCTACGACATGAGCTGGGTCCGTCAGGCTCCAGGCAAGGGACTGGAGTGGGTGGGACTCATTAGTTATGATGGTAACACATACTACGCGACCTCCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTACCTGCAAATGTCTAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCTAGAAGTCTTTATGCTGGTCCTAATGCTGGTATCGGACCGTTTAACATCTGGGGCCAAGGTACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTG TCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 315; SEQ ID NO: 316; and SEQ ID NO: 317 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 111 or the light chain sequence of SEQ ID NO: 112.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 318; SEQ ID NO: 319; and SEQ ID NO: 320 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 113 or the heavy chain sequence of SEQ ID NO: 114.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 311 encoding the light chain variable sequenceof SEQ ID NO: 111; the polynucleotide SEQ ID NO: 312 encoding the lightchain sequence of SEQ ID NO: 112; the polynucleotide SEQ ID NO: 313encoding the heavy chain variable sequence of SEQ ID NO: 113; thepolynucleotide SEQ ID NO: 314 encoding the heavy chain sequence of SEQID NO: 114; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 315; SEQ ID NO: 316; and SEQ ID NO: 317) of thelight chain variable sequence of SEQ ID NO: 111 or the light chainsequence of SEQ ID NO: 112; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 318; SEQ ID NO: 319; andSEQ ID NO: 320) of the heavy chain variable sequence of SEQ ID NO: 113or the heavy chain sequence of SEQ ID NO: 114.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab12, the polynucleotidesencoding the full length Ab12 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 312 encoding the light chainsequence of SEQ ID NO: 112 and the polynucleotide SEQ ID NO: 314encoding the heavy chain sequence of SEQ ID NO: 114.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as yeast cells such as theyeast Pichia. Suitable Pichia species include, but are not limited to,Pichia pastoris. In one embodiment of the invention described herein(infra), Fab fragments may be produced by enzymatic digestion (e.g.,papain) of Ab12 following expression of the full-length polynucleotidesin a suitable host. In another embodiment of the invention, anti-NGFantibodies such as Ab12 or Fab fragments thereof may be produced viaexpression of Ab12 polynucleotides in mammalian cells such as CHO, NSOor HEK 293 cells, fungal, insect, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab13

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab13 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab13 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 121:

(SEQ ID NO: 321) GCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCGGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACGTGCAGTGTGACGCTGCTGCCACTTACTACTGTGCAGGCGGTTATACCAGTAGTAGTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:122:

(SEQ ID NO: 322) GCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCGGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACGTGCAGTGTGACGCTGCTGCCACTTACTACTGTGCAGGCGGTTATACCAGTAGTAGTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGG AGAGTGTTA.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 123:

(SEQ ID NO: 323) CAGTCGGTGGAGGCGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTACCTACTGGATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGACATTTATTTTAGTAATGAAGAAACAAACTACGCGAGCTGGGCGAAAGGCCGATTTACCATCTCCAAAACCTCGACCACGGTGGATCTGAATGTCATCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGTTCTCCTGATGTTGATATTGGTATAGATATGTGGGGCCCGGGCACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:124:

(SEQ ID NO: 324) CAGTCGGTGGAGGCGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTACCTACTGGATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGACATTTATTTTAGTAATGAAGAAACAAACTACGCGAGCTGGGCGAAAGGCCGATTTACCATCTCCAAAACCTCGACCACGGTGGATCTGAATGTCATCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGTTCTCCTGATGTTGATATTGGTATAGATATGTGGGGCCCGGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCAcCCTCCTCCaAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 325; SEQ ID NO: 326; and SEQ ID NO: 327 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 121 or the light chain sequence of SEQ ID NO: 122.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 328; SEQ ID NO: 329; and SEQ ID NO: 330 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 123 or the heavy chain sequence of SEQ ID NO: 124.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 321 encoding the light chain variable sequenceof SEQ ID NO: 121; the polynucleotide SEQ ID NO: 322 encoding the lightchain sequence of SEQ ID NO: 122; the polynucleotide SEQ ID NO: 323encoding the heavy chain variable sequence of SEQ ID NO: 123; thepolynucleotide SEQ ID NO: 324 encoding the heavy chain sequence of SEQID NO: 124; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 325; SEQ ID NO: 326; and SEQ ID NO: 327) of thelight chain variable sequence of SEQ ID NO: 121 or the light chainsequence of SEQ ID NO: 122; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 328; SEQ ID NO: 329; andSEQ ID NO: 330) of the heavy chain variable sequence of SEQ ID NO: 123or the heavy chain sequence of SEQ ID NO: 124.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab13, the polynucleotidesencoding the full length Ab13 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 322 encoding the light chainsequence of SEQ ID NO: 122 and the polynucleotide SEQ ID NO: 324encoding the heavy chain sequence of SEQ ID NO: 124.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as yeast cells such as theyeast Pichia. Suitable Pichia species include, but are not limited to,Pichia pastoris. In one embodiment of the invention described herein(infra), Fab fragments may be produced by enzymatic digestion (e.g.,papain) of Ab13 following expression of the full-length polynucleotidesin a suitable host. In another embodiment of the invention, anti-NGFantibodies such as Ab13 or Fab fragments thereof may be produced viaexpression of Ab13 polynucleotides in mammalian cells such as CHO, NSOor HEK 293 cells, fungal, insect, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab14

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab14 polypeptides having bindingspecificity to NGF which further inhibit the association of NGF withTrkA and the association of NGF with p75, in methods of treating pain inan individual comprising administering to said individual antibody Ab14polypeptides. The invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to NGF. In oneembodiment of the invention, polynucleotides of the invention comprise,or alternatively consist of, the following polynucleotide sequenceencoding the variable light chain polypeptide sequence of SEQ ID NO:131:

(SEQ ID NO: 331) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATACCAGTAGTAGTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:132:

(SEQ ID NO: 332) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATACCAGTAGTAGTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGA GAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 133:

(SEQ ID NO: 333) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTACCTACTGGATGAGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGACATTTACTTTAGTAATGAAGAAACAAACTACGCGAGCAGCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGTTCTCCTGATGTTGATATTGGTATAGATATGTGGGGCCCAGGGACCCTCGT CACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:134:

(SEQ ID NO: 334) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTACCTACTGGATGAGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGACATTTACTTTAGTAATGAAGAAACAAACTACGCGAGCAGCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGTTCTCCTGATGTTGATATTGGTATAGATATGTGGGGCCCAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 335; SEQ ID NO: 336; and SEQ ID NO: 337 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 131 or the light chain sequence of SEQ ID NO: 132.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 338; SEQ ID NO: 339; and SEQ ID NO: 340 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 133 or the heavy chain sequence of SEQ ID NO: 134.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 331 encoding the light chain variable sequenceof SEQ ID NO: 131; the polynucleotide SEQ ID NO: 332 encoding the lightchain sequence of SEQ ID NO: 132; the polynucleotide SEQ ID NO: 333encoding the heavy chain variable sequence of SEQ ID NO: 133; thepolynucleotide SEQ ID NO: 334 encoding the heavy chain sequence of SEQID NO: 134; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 335; SEQ ID NO: 336; and SEQ ID NO: 337) of thelight chain variable sequence of SEQ ID NO: 131 or the light chainsequence of SEQ ID NO: 132; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 338; SEQ ID NO: 339; andSEQ ID NO: 340) of the heavy chain variable sequence of SEQ ID NO: 133or the heavy chain sequence of SEQ ID NO: 134.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab14, the polynucleotidesencoding the full length Ab14 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 332 encoding the light chainsequence of SEQ ID NO: 132 and the polynucleotide SEQ ID NO: 334encoding the heavy chain sequence of SEQ ID NO: 134.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as bacterial or yeast cellssuch as the yeast Pichia. Suitable Pichia species include, but are notlimited to, Pichia pastoris. In one embodiment of the inventiondescribed herein (infra), Fab fragments may be produced by enzymaticdigestion (e.g., papain) of Ab14 following expression of the full-lengthpolynucleotides in a suitable host. In another embodiment of theinvention, anti-NGF antibodies such as Ab14 or Fab fragments thereof maybe produced via expression of Ab14 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab15

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab15 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA and donot appreciably affect the association of NGF with p75, in methods oftreating pain in an individual comprising administering to saidindividual antibody Ab15 polypeptides. The invention is further directedto polynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 141:

(SEQ ID NO: 341) GCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCAGCTGTGGGAGACACAGTCACCATCAAGTGCCAGTCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGATGCATCCAATCTGCCATCTGGGGTCCCATCACGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGATTATGATGATGATACTGATAATGGTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:142:

(SEQ ID NO: 342) GCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCAGCTGTGGGAGACACAGTCACCATCAAGTGCCAGTCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGATGCATCCAATCTGCCATCTGGGGTCCCATCACGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGATTATGATGATGATACTGATAATGGTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 143:

(SEQ ID NO: 343) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGACCTCAGTAGCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGAATCATTTGGAGTGGTGGCACCTACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGCAAATCACCAGTCCGACAACCGAGGACGCGGCCACCTATTTCTGTGCCGCAGGTGGTGGTAGTATTTATGATGTTTGGGGCCCGGGCACCCTGGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:144:

(SEQ ID NO: 344) CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGACCTCAGTAGCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGAATCATTTGGAGTGGTGGCACCTACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGCAAATCACCAGTCCGACAACCGAGGACGCGGCCACCTATTTCTGTGCCGCAGGTGGTGGTAGTATTTATGATGTTTGGGGCCCGGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 345; SEQ ID NO: 346; and SEQ ID NO: 347 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 141 or the light chain sequence of SEQ ID NO: 142.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 348; SEQ ID NO: 349; and SEQ ID NO: 350 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 143 or the heavy chain sequence of SEQ ID NO: 144.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 341 encoding the light chain variable sequenceof SEQ ID NO: 141; the polynucleotide SEQ ID NO: 342 encoding the lightchain sequence of SEQ ID NO: 142; the polynucleotide SEQ ID NO: 343encoding the heavy chain variable sequence of SEQ ID NO: 143; thepolynucleotide SEQ ID NO: 344 encoding the heavy chain sequence of SEQID NO: 144; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 345; SEQ ID NO: 346; and SEQ ID NO: 347) of thelight chain variable sequence of SEQ ID NO: 141 or the light chainsequence of SEQ ID NO: 142; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 348; SEQ ID NO: 349; andSEQ ID NO: 350) of the heavy chain variable sequence of SEQ ID NO: 143or the heavy chain sequence of SEQ ID NO: 144.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab15, the polynucleotidesencoding the full length Ab15 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 342 encoding the light chainsequence of SEQ ID NO: 142 and the polynucleotide SEQ ID NO: 344encoding the heavy chain sequence of SEQ ID NO: 144.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab15 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab15 or Fab fragments thereofmay be produced via expression of Ab15 polynucleotides in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab16

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab16 polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF which inhibit the association of NGF with TrkA and donot appreciably affect the association of NGF with p75, in methods oftreating pain in an individual comprising administering to saidindividual antibody Ab16 polypeptides. The invention is further directedto polynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 151:

(SEQ ID NO: 351) GCCCTGGTGATGACCCAGACTCCATCCTCCACGTCTGAACCAGTGGGAGGCACAGTCACCATCAATTGCCAGGCTAGTCAGAATATTGGTAACGACCTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCGAGCTCCTAATCTATTCTACATCCAAACTGGCAACTGGGGTCCCAAAGCGGTTCAGTGGCAGCAGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTCTAGGTGTTTATAGTTATATTAGTGATGATGGTAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:152:

(SEQ ID NO: 352) GCCCTGGTGATGACCCAGACTCCATCCTCCACGTCTGAACCAGTGGGAGGCACAGTCACCATCAATTGCCAGGCTAGTCAGAATATTGGTAACGACCTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCGAGCTCCTAATCTATTCTACATCCAAACTGGCAACTGGGGTCCCAAAGCGGTTCAGTGGCAGCAGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTCTAGGTGTTTATAGTTATATTAGTGATGATGGTAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 153:

(SEQ ID NO: 353) CAGTCGGTGGAGGAGTTCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACCGTCTCTGGATTCTCCCTCAATAACTATGCAATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGGATCATTGGTAGTATTGGTACCACATACTACGCGAGCTGGGCGAAAGGCCGATTCTTCATCTCCAAAACCTCGACCACTGTGGATCTGAAAATCATTAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGATGCTGGCGTTACTGTTGATGGTTATGGCTACTACTTTAACATCTGGGGCCCAGGCACCCTCGTCAC CGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:154:

(SEQ ID NO: 354) CAGTCGGTGGAGGAGTTCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACCGTCTCTGGATTCTCCCTCAATAACTATGCAATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGGATCATTGGTAGTATTGGTACCACATACTACGCGAGCTGGGCGAAAGGCCGATTCTTCATCTCCAAAACCTCGACCACTGTGGATCTGAAAATCATTAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGATGCTGGCGTTACTGTTGATGGTTATGGCTACTACTTTAACATCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 355; SEQ ID NO: 356; and SEQ ID NO: 357 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 151 or the light chain sequence of SEQ ID NO: 152.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 358; SEQ ID NO: 359; and SEQ ID NO: 360 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 153 or the heavy chain sequence of SEQ ID NO: 154.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 351 encoding the light chain variable sequenceof SEQ ID NO: 151; the polynucleotide SEQ ID NO: 352 encoding the lightchain sequence of SEQ ID NO: 152; the polynucleotide SEQ ID NO: 353encoding the heavy chain variable sequence of SEQ ID NO: 153; thepolynucleotide SEQ ID NO: 354 encoding the heavy chain sequence of SEQID NO: 154; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 355; SEQ ID NO: 356; and SEQ ID NO: 357) of thelight chain variable sequence of SEQ ID NO: 151 or the light chainsequence of SEQ ID NO: 152; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 358; SEQ ID NO: 359; andSEQ ID NO: 360) of the heavy chain variable sequence of SEQ ID NO: 153or the heavy chain sequence of SEQ ID NO: 154.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab16, the polynucleotidesencoding the full length Ab16 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 352 encoding the light chainsequence of SEQ ID NO: 152 and the polynucleotide SEQ ID NO: 354encoding the heavy chain sequence of SEQ ID NO: 154.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as yeast cells such as theyeast Pichia. Suitable Pichia species include, but are not limited to,Pichia pastoris. In one embodiment of the invention described herein(infra), Fab fragments may be produced by enzymatic digestion (e.g.,papain) of Ab16 following expression of the full-length polynucleotidesin a suitable host. In another embodiment of the invention, anti-NGFantibodies such as Ab16 or Fab fragments thereof may be produced viaexpression of Ab16 polynucleotides in mammalian cells such as CHO, NSOor HEK 293 cells, fungal, insect, or microbial systems such as yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab17

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab17 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab17 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 161:

(SEQ ID NO: 361) GCCATCGAAATGACCCAGACTCCATTCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGACCATTAGCAACTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTCAACAGGGTTATACTATCAGTAATGTTGATAACAATGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:162:

(SEQ ID NO: 362) GCCATCGAAATGACCCAGACTCCATTCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGACCATTAGCAACTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTCAACAGGGTTATACTATCAGTAATGTTGATAACAATGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 163:

(SEQ ID NO: 363) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGGGATCCCTGACACTCACCTGCGCAGCCTCTGGATTCTCCCTCACTGGCTACAACTTGGTCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGATTCATTAGTTATGGTGATACCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGACTCTGACGATCACCGATCTGCAACCTTCAGACACGGGCACCTATTTCTGTGCCAGAGAGACTGCTAATACTTATGATTATGGCATCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:164:

(SEQ ID NO: 364) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGGGATCCCTGACACTCACCTGCGCAGCCTCTGGATTCTCCCTCACTGGCTACAACTTGGTCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGATTCATTAGTTATGGTGATACCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGACTCTGACGATCACCGATCTGCAACCTTCAGACACGGGCACCTATTTCTGTGCCAGAGAGACTGCTAATACTTATGATTATGGCATCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 365; SEQ ID NO: 366; and SEQ ID NO: 367 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 161 or the light chain sequence of SEQ ID NO: 162.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 368; SEQ ID NO: 369; and SEQ ID NO: 370 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 163 or the heavy chain sequence of SEQ ID NO: 164.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 361 encoding the light chain variable sequenceof SEQ ID NO: 161; the polynucleotide SEQ ID NO: 362 encoding the lightchain sequence of SEQ ID NO: 162; the polynucleotide SEQ ID NO: 363encoding the heavy chain variable sequence of SEQ ID NO: 163; thepolynucleotide SEQ ID NO: 364 encoding the heavy chain sequence of SEQID NO: 164; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 365; SEQ ID NO: 366; and SEQ ID NO: 367) of thelight chain variable sequence of SEQ ID NO: 161 or the light chainsequence of SEQ ID NO: 162; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 368; SEQ ID NO: 369; andSEQ ID NO: 370) of the heavy chain variable sequence of SEQ ID NO: 163or the heavy chain sequence of SEQ ID NO: 164.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab17, the polynucleotidesencoding the full length Ab17 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 362 encoding the light chainsequence of SEQ ID NO: 162 and the polynucleotide SEQ ID NO: 364encoding the heavy chain sequence of SEQ ID NO: 164.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as bacterial or yeast cellssuch as the yeast Pichia. Suitable Pichia species include, but are notlimited to, Pichia pastoris. In one embodiment of the inventiondescribed herein (infra), Fab fragments may be produced by enzymaticdigestion (e.g., papain) of Ab17 following expression of the full-lengthpolynucleotides in a suitable host. In another embodiment of theinvention, anti-NGF antibodies such as Ab17 or Fab fragments thereof maybe produced via expression of Ab17 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab18

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab18 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab18 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 171:

(SEQ ID NO: 371) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGTCAGGCTAGTCAGACCATTAGCAACTACTTAGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGAACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGTCAACAGGGTTATACTATCAGTAATGTTGATAACAATGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:172:

(SEQ ID NO: 372) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGTCAGGCTAGTCAGACCATTAGCAACTACTTAGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGAACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGTCAACAGGGTTATACTATCAGTAATGTTGATAACAATGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 173:

(SEQ ID NO: 373) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTGGCTACAACTTGGTCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGATTCATTAGTTATGGTGATACCACATACTACGCTAGCTCTGCTAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGAGACTGCTAATACTTATGATTATGGCATCTGGGGCCAAGGGACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:174:

(SEQ ID NO: 374) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTGGCTACAACTTGGTCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGATTCATTAGTTATGGTGATACCACATACTACGCTAGCTCTGCTAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGAGACTGCTAATACTTATGATTATGGCATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 375; SEQ ID NO: 376; and SEQ ID NO: 377 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 171 or the light chain sequence of SEQ ID NO: 172.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 378; SEQ ID NO: 379; and SEQ ID NO: 380 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 173 or the heavy chain sequence of SEQ ID NO: 174.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 371 encoding the light chain variable sequenceof SEQ ID NO: 171; the polynucleotide SEQ ID NO: 372 encoding the lightchain sequence of SEQ ID NO: 172; the polynucleotide SEQ ID NO: 373encoding the heavy chain variable sequence of SEQ ID NO: 173; thepolynucleotide SEQ ID NO: 374 encoding the heavy chain sequence of SEQID NO: 174; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 375; SEQ ID NO: 376; and SEQ ID NO: 377) of thelight chain variable sequence of SEQ ID NO: 171 or the light chainsequence of SEQ ID NO: 172; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 378; SEQ ID NO: 379; andSEQ ID NO: 380) of the heavy chain variable sequence of SEQ ID NO: 173or the heavy chain sequence of SEQ ID NO: 174.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments for treatment orprevention of pain and pain associated conditions having bindingspecificity for NGF. With respect to antibody Ab18, the polynucleotidesencoding the full length Ab18 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 372 encoding the light chainsequence of SEQ ID NO: 172 and the polynucleotide SEQ ID NO: 374encoding the heavy chain sequence of SEQ ID NO: 174.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab18 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab18 or Fab fragments thereofmay be produced via expression of Ab18 polynucleotides in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as bacterial oryeast cells (for example diploid yeast such as diploid Pichia) and otheryeast strains. Suitable Pichia species include, but are not limited to,Pichia pastoris.

Antibody Ab19

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab19 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab19 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 181:

(SEQ ID NO: 381) GCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTATTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCTTCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACGTGCAGTGTGACGCTGCTGCCACTTACTACTGTGCAGGCGGTTATAGTAGTAGTAGTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:182:

(SEQ ID NO: 382) GCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTATTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCTTCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACGTGCAGTGTGACGCTGCTGCCACTTACTACTGTGCAGGCGGTTATAGTAGTAGTAGTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 183:

(SEQ ID NO: 383) CAGTCGGTGGAGGCGTCCGGGGGTCGTCTGGTCATGCCTGGAGGATCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTACCTACTGGATGTCCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGACATTTATTTTAGTAATGAGGAAACAAACTACGCGACCTGGGCGAAAGGCCGATTTACCATCTCCAAAACCTCGACCACGGTGGATCTGAATGTCATCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCAAGAGGTTCTCCTGATGTTGAGATTGCTATAGATATGTGGGGCCAGGGCACCCTCGTCACCGTCTCGAG C.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:184:

(SEQ ID NO: 384) CAGTCGGTGGAGGCGTCCGGGGGTCGTCTGGTCATGCCTGGAGGATCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTACCTACTGGATGTCCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGACATTTATTTTAGTAATGAGGAAACAAACTACGCGACCTGGGCGAAAGGCCGATTTACCATCTCCAAAACCTCGACCACGGTGGATCTGAATGTCATCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCAAGAGGTTCTCCTGATGTTGAGATTGCTATAGATATGTGGGGCCAGGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCAcCCTCCTCCaAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 385; SEQ ID NO: 386; and SEQ ID NO: 387 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 181 or the light chain sequence of SEQ ID NO: 182.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 388; SEQ ID NO: 389; and SEQ ID NO: 390 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 183 or the heavy chain sequence of SEQ ID NO: 184.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 381 encoding the light chain variable sequenceof SEQ ID NO: 181; the polynucleotide SEQ ID NO: 382 encoding the lightchain sequence of SEQ ID NO: 182; the polynucleotide SEQ ID NO: 383encoding the heavy chain variable sequence of SEQ ID NO: 183; thepolynucleotide SEQ ID NO: 384 encoding the heavy chain sequence of SEQID NO: 184; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 385; SEQ ID NO: 386; and SEQ ID NO: 387) of thelight chain variable sequence of SEQ ID NO: 181 or the light chainsequence of SEQ ID NO: 182; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 388; SEQ ID NO: 389; andSEQ ID NO: 390) of the heavy chain variable sequence of SEQ ID NO: 183or the heavy chain sequence of SEQ ID NO: 184.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab19, the polynucleotidesencoding the full length Ab19 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 382 encoding the light chainsequence of SEQ ID NO: 182 and the polynucleotide SEQ ID NO: 384encoding the heavy chain sequence of SEQ ID NO: 184.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab19 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab19 or Fab fragments thereofmay be produced via expression of Ab19 polynucleotides in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab20

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab20 polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab20 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 191:

(SEQ ID NO: 391) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATACCAGTAGTAGTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:192:

(SEQ ID NO: 392) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAATGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGTCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATACCAGTAGTAGTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 193:

(SEQ ID NO: 393) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTACCTACTGGATGAGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGACATTTACTTTAGTAATGAAGAAACAAACTACGCGACCAGCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGTTCTCCTGATGTTGAGATTGCTATAGATATGTGGGGCCAAGGGACCCTCGTCAC CGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:194:

(SEQ ID NO: 394) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTACCTACTGGATGAGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGACATTTACTTTAGTAATGAAGAAACAAACTACGCGACCAGCGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGTTCTCCTGATGTTGAGATTGCTATAGATATGTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 395; SEQ ID NO: 396; and SEQ ID NO: 397 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 191 or the light chain sequence of SEQ ID NO: 192.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 398; SEQ ID NO: 399; and SEQ ID NO: 400 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 193 or the heavy chain sequence of SEQ ID NO: 194.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 391 encoding the light chain variable sequenceof SEQ ID NO: 191; the polynucleotide SEQ ID NO: 392 encoding the lightchain sequence of SEQ ID NO: 192; the polynucleotide SEQ ID NO: 393encoding the heavy chain variable sequence of SEQ ID NO: 193; thepolynucleotide SEQ ID NO: 394 encoding the heavy chain sequence of SEQID NO: 194; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 395; SEQ ID NO: 396; and SEQ ID NO: 397) of thelight chain variable sequence of SEQ ID NO: 191 or the light chainsequence of SEQ ID NO: 192; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 398; SEQ ID NO: 399; andSEQ ID NO: 400) of the heavy chain variable sequence of SEQ ID NO: 193or the heavy chain sequence of SEQ ID NO: 194.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab20, the polynucleotidesencoding the full length Ab20 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 392 encoding the light chainsequence of SEQ ID NO: 192 and the polynucleotide SEQ ID NO: 394encoding the heavy chain sequence of SEQ ID NO: 194.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab20 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-NGF antibodies such as Ab20 or Fab fragments thereofmay be produced via expression of Ab20 polynucleotides in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, plant cell,transgenic plant or animal, or microbial systems such as yeast cells(for example diploid yeast such as diploid Pichia) and other yeaststrains. Suitable Pichia species include, but are not limited to, Pichiapastoris.

Antibody Ab21

The invention is further directed to the use of polynucleotides setforth below to produce antibody Ab21 polypeptides having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab21 polypeptides. The invention is further directed topolynucleotides encoding antibody polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, polynucleotidesof the invention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable light chain polypeptidesequence of SEQ ID NO: 51:

(SEQ ID NO: 251) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTTACAGCAATCTTGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:401:

(SEQ ID NO: 403) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTTACAGCAATCTTGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 53:

(SEQ ID NO: 253) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAACTATGCAGTGGGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTCTGCAAGAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGATATGGCCGTAGTGTTGCTTATTACGTCTTTAACATCTGGGGCCCAGGGACCCTCGT CACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:402:

(SEQ ID NO: 404) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAACTATGCAGTGGGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTCTGCAAGAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGATATGGCCGTAGTGTTGCTTACTACGTCTTTAACATCTGGGGCCCAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACGCGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 51 or the light chain sequence of SEQ ID NO: 401.

In a further embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 258; SEQ ID NO: 259; and SEQ ID NO: 260 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 53 or the heavy chain sequence of SEQ ID NO: 402.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 251 encoding the light chain variable sequenceof SEQ ID NO: 51; the polynucleotide SEQ ID NO: 403 encoding the lightchain sequence of SEQ ID NO: 401; the polynucleotide SEQ ID NO: 253encoding the heavy chain variable sequence of SEQ ID NO: 53; thepolynucleotide SEQ ID NO: 404 encoding the heavy chain sequence of SEQID NO: 402; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257) of thelight chain variable sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 401; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 258; SEQ ID NO: 259; andSEQ ID NO: 260) of the heavy chain variable sequence of SEQ ID NO: 53 orthe heavy chain sequence of SEQ ID NO: 402.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody Ab21, the polynucleotidesencoding the full length Ab21 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 403 encoding the light chainsequence of SEQ ID NO: 401 and the polynucleotide SEQ ID NO: 404encoding the heavy chain sequence of SEQ ID NO: 402.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, plant cell, transgenicplant or animal, or microbial systems such as bacterial or yeast cellssuch as the yeast Pichia. Suitable Pichia species include, but are notlimited to, Pichia pastoris. In one embodiment of the inventiondescribed herein (infra), Fab fragments may be produced by enzymaticdigestion (e.g., papain) of Ab21 following expression of the full-lengthpolynucleotides in a suitable host. In another embodiment of theinvention, anti-NGF antibodies such as Ab21 or Fab fragments thereof maybe produced via expression of Ab21 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Fragment Fab2

The invention is further directed to the use of polynucleotides setforth below to produce antibody fragment Fab2 polypeptides for treatmentor prevention of pain and pain associated conditions having bindingspecificity to NGF which inhibit the association of NGF with TrkA andfurther inhibit the association of NGF with p75, in methods of treatingpain in an individual comprising administering to said individualantibody Ab1 polypeptides. The invention is further directed topolynucleotides encoding antibody fragment polypeptides for treatment orprevention of pain and pain associated conditions having bindingspecificity to NGF. In one embodiment of the invention, Fabpolynucleotides of the invention comprise, or alternatively consist of,the following polynucleotide sequence encoding the light chainpolypeptide sequence of SEQ ID NO: 407:

(SEQ ID NO: 409) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTTACAGCAATCTTGCCTGGTATCAGCAGAAACCAGGAAAAGCCCCTAAGCTCCTGATCTATGATGCATCCACTCTGGAATCTGGAGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTACTACTGCCAACAGGGTTTTACTGTTAGTGATATTGATAATGCTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTG TTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:408:

(SEQ ID NO: 410) GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCGTCAGTAACTATGCAGTGGGCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAATCATTGGTCGTAATGGTAACACATGGTACGCGAGCTCTGCAAGAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGGATATGGCCGTAGTGTTGCTTACTACGTCTTTAACATCTGGGGCCCAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACGCGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACTAG.

In a further embodiment of the invention, polynucleotides encoding Fabantibody fragments having binding specificity to NGF comprise one ormore of the polynucleotide sequences of SEQ ID NO: 255; SEQ ID NO: 256;and SEQ ID NO: 257 which correspond to polynucleotides encoding thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain variable sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 409.

In a further embodiment of the invention, polynucleotides encoding Fabantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise one ormore of the polynucleotide sequences of SEQ ID NO: 258; SEQ ID NO: 259;and SEQ ID NO: 260 which correspond to polynucleotides encoding thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain variable sequence of SEQ ID NO: 53 or the heavy chainsequence of SEQ ID NO: 410.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragments fortreatment or prevention of pain and pain associated conditions describedherein. In one embodiment of the invention, polynucleotides encodingantibody fragments for treatment or prevention of pain and painassociated conditions having binding specificity to NGF comprise, oralternatively consist of, one, two, three or more, including all of thefollowing polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 251 encoding the light chain variable sequenceof SEQ ID NO: 51; the polynucleotide SEQ ID NO: 409 encoding the lightchain sequence of SEQ ID NO: 407; the polynucleotide SEQ ID NO: 253encoding the heavy chain variable sequence of SEQ ID NO: 53; thepolynucleotide SEQ ID NO: 410 encoding the heavy chain sequence of SEQID NO: 408; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257) of thelight chain variable sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 407; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 258; SEQ ID NO: 259; andSEQ ID NO: 260) of the heavy chain variable sequence of SEQ ID NO: 53 orthe heavy chain sequence of SEQ ID NO: 408.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for NGF. With respect to antibody fragment Fab2, thepolynucleotides encoding the Fab fragment include the polynucleotide SEQID NO: 409 encoding the light chain sequence of SEQ ID NO: 407 and thepolynucleotide SEQ ID NO: 410 encoding the heavy chain sequence of SEQID NO: 408.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be producedvia expression of Fab2 polynucleotides in mammalian cells such as CHO,NSO or HEK 293 cells, fungal, insect, plant cell, transgenic plant oranimal, or microbial systems such as bacterial or yeast cells (forexample diploid yeast such as diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In one embodiment, the invention is directed to an isolatedpolynucleotide comprising a polynucleotide encoding an anti-NGF V_(H)antibody amino acid sequence selected from SEQ ID NO: 3, 13, 23, 33, 43,53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, or402, or encoding a variant thereof wherein at least one frameworkresidue (FR residue) has been substituted with an amino acid present atthe corresponding position in a rabbit anti-NGF antibody V_(H)polypeptide or a conservative amino acid substitution.

In another embodiment, the invention is directed to an isolatedpolynucleotide comprising the polynucleotide sequence encoding ananti-NGF V_(L) antibody amino acid sequence of 1, 11, 21, 31, 41, 51,61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, or401, or encoding a variant thereof wherein at least one frameworkresidue (FR residue) has been substituted with an amino acid present atthe corresponding position in a rabbit anti-NGF antibody V_(L)polypeptide or a conservative amino acid substitution.

In yet another embodiment, the invention is directed to one or moreheterologous polynucleotides comprising a sequence encoding thepolypeptides contained in SEQ ID NO:1 and SEQ ID NO:3; SEQ ID NO:11 andSEQ ID NO:13; SEQ ID NO:21 and SEQ ID NO:23; SEQ ID NO:31 and SEQ IDNO:33; SEQ ID NO:411 and SEQ ID NO:43; SEQ ID NO:51 and SEQ ID NO:53,SEQ ID NO:61 and SEQ ID NO:63; SEQ ID NO:71 and SEQ ID NO:73; SEQ IDNO:81 and SEQ ID NO:83; SEQ ID NO:91 and SEQ ID NO:93; SEQ ID NO:101 andSEQ ID NO:103; SEQ ID NO:111 and SEQ ID NO:113; SEQ ID NO:121 and SEQ IDNO:123; SEQ ID NO:131 and SEQ ID NO:133; SEQ ID NO:141 and SEQ IDNO:143; SEQ ID NO:151 and SEQ ID NO:153; SEQ ID NO:161 and SEQ IDNO:163; SEQ ID NO:171 and SEQ ID NO:173; SEQ ID NO:181 and SEQ IDNO:183; SEQ ID NO:191 and SEQ ID NO:193; or SEQ ID NO:401 and SEQ IDNO:403.

In another embodiment, the invention is directed to an isolatedpolynucleotide that expresses a polypeptide containing at least one CDRpolypeptide derived from an anti-NGF antibody wherein said expressedpolypeptide alone specifically binds NGF or specifically binds NGF whenexpressed in association with another polynucleotide sequence thatexpresses a polypeptide containing at least one CDR polypeptide derivedfrom an anti-NGF antibody for treatment or prevention of pain and painassociated conditions wherein said at least one CDR is selected fromthose contained in the V_(L) or V_(H) polypeptides of SEQ ID NO: 1, 3,11, 13, 21, 23, 31, 33, 41, 43, 51, 53, 61, 63, 71, 73, 81, 83, 91, 93,101, 103, 111, 113, 121, 123, 131, 133, 141, 143, 151, 153, 161, 163,171, 173, 181, 183, 191, 193, 401 or SEQ ID NO:403.

Host cells and vectors comprising said polynucleotides are alsocontemplated.

The invention further contemplates vectors comprising the polynucleotidesequences encoding the variable heavy and light chain polypeptidesequences, as well as the individual complementarity-determining regions(CDRs, or hypervariable regions), as set forth herein, as well as hostcells comprising said vector sequences. In one embodiment of theinvention, the host cell is a yeast cell. In another embodiment of theinvention, the yeast host cell belongs to the genus Pichia.

Anti-NGF Activity

The anti-NGF activity of the anti-NGF antibodies of the presentinvention, and fragments thereof having binding specificity to NGF,preferably which inhibit the association of NGF with TrkA and/or p75,may also be described by their strength of binding or their affinity forNGF. In one embodiment of the invention, the anti-NGF antibodies of thepresent invention, and fragments thereof having binding specificity toNGF, bind to NGF with a dissociation constant (K_(D)) of less than orequal to 5×10⁻⁷ M, 10⁻⁷ M, 5×10⁻⁸ M, 10⁻⁸M, 5×10⁻⁹ M, 10⁻⁹ M, 5×10⁻¹⁰ M,10⁻¹⁰ M, 5×10⁻¹¹ M, 10⁻¹¹ M, 5×10⁻¹² M, 10⁻¹² M, 5×10⁻¹³ M, or 10⁻¹³ M.Preferably, the anti-NGF antibodies and fragments thereof bind NGF witha dissociation constant of less than or equal to 5×10⁻¹⁰ M. In anotherembodiment of the invention, the anti-NGF antibodies of the presentinvention, and fragments thereof having binding specificity to NGF, bindto a linear or conformational NGF epitope.

In another embodiment of the invention, the anti-NGF activity of theanti-NGF antibodies of the present invention, and fragments thereofhaving binding specificity to NGF, bind to NGF with an off-rate of lessthan or equal to 10⁻⁴ S⁻¹, 5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹, 5×10⁻⁶ S⁻¹, 10⁻⁶ S⁻¹,5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹.

In a further embodiment of the invention, the anti-NGF activity of theanti-NGF antibodies of the present invention, and fragments thereofhaving binding specificity to NGF, exhibit anti-NGF activity bypreventing, ameliorating or reducing the symptoms of, or alternativelytreating, diseases and disorders associated with NGF, and preferredpain-related diseases and disorders, and selectively inhibit theassociation of NGF with TrkA and/or p75. Non-limiting examples ofdiseases and disorders associated with NGF are set forth infra.

The invention is especially directed to methods of treating pain usingchimeric or humanized antibodies and fragments thereof (including Fabfragments) capable of binding to NGF which inhibit the association ofNGF with TrkA and/or p75. However, the invention further encompassesusing chimeric or humanized antibodies and fragments thereof (includingFab fragments) capable of binding and/or inhibiting the biologicalactivities mediated by the binding of NGF that do not inhibit theassociation of NGF with the p75 and TrkA receptors. In another preferredembodiment of the invention, full length antibodies and Fab fragmentsthereof are capable of significantly reducing pain in vivo in murinemodels, as measured by Gait analysis (as described in the examplesherein) that further which inhibit the association of NGF with TrkAand/or p75.

B-cell Screening and Isolation

In one embodiment, the present invention contemplates the preparationand isolation of a clonal population of antigen-specific B cells thatmay be used for isolating at least one NGF antigen-specific cell, whichcan be used to produce a monoclonal antibody against NGF, which isspecific to a desired NGF antigen, or a nucleic acid sequencecorresponding to such an antibody. Methods of preparing and isolatingsaid clonal population of antigen-specific B cells are taught, forexample, in U.S. patent publication no. US 2007/0269868 toCarvalho-Jensen et al., the disclosure of which is herein incorporatedby reference in its entirety. Methods of preparing and isolating saidclonal population of antigen-specific B cells are also taught herein inthe examples. Methods of “enriching” a cell population by size ordensity are known in the art. See, e.g., U.S. Pat. No. 5,627,052. Thesesteps can be used in addition to enriching the cell population byantigen-specificity.

Methods of Humanizing Antibodies

In another embodiment, the present invention contemplates methods forhumanizing antibody heavy and light chains. Methods for humanizingantibody heavy and light chains which may be applied to anti-NGFantibodies are taught, for example, in U.S. patent applicationpublication no. US 2009/0022659 to Olson et al., and in U.S. patentapplication publication no. US 2009/0028784 to Garcia-Martinez et al.,the disclosures of each of which are herein incorporated by reference intheir entireties.

Methods of Producing Antibodies and Fragments Thereof

In another embodiment, the present invention contemplates methods forproducing anti-NGF antibodies and fragments thereof. Methods forproducing anti-NGF antibodies and fragments thereof secreted frompolyploidal, preferably diploid or tetraploid strains of matingcompetent yeast are taught, for example, in U.S. patent applicationpublication no. US 2009/0022659 to Olson et al., and in U.S. patentapplication publication no. US 2009/0028784 to Garcia-Martinez et al.,the disclosures of each of which are herein incorporated by reference intheir entireties.

Other methods of producing antibodies are well known to those ofordinary skill in the art. For example, methods of producing chimericantibodies are now well known in the art (See, for example, U.S. Pat.No. 4,816,567 to Cabilly et al.; Morrison et al., P.N.A.S. USA,81:8651-55 (1984); Neuberger, M. S. et al., Nature, 314:268-270 (1985);Boulianne, G. L. et al., Nature, 312:643-46 (1984), the disclosures ofeach of which are herein incorporated by reference in their entireties).

Likewise, other methods of producing humanized antibodies are now wellknown in the art (See, for example, U.S. Pat. Nos. 5,530,101, 5,585,089,5,693,762, and 6,180,370 to Queen et al; U.S. Pat. Nos. 5,225,539 and6,548,640 to Winter; U.S. Pat. Nos. 6,054,297, 6,407,213 and 6,639,055to Carter et al; U.S. Pat. No. 6,632,927 to Adair; Jones, P. T. et al,Nature, 321:522-525 (1986); Reichmann, L., et al, Nature, 332:323-327(1988); Verhoeyen, M, et al, Science, 239:1534-36 (1988), thedisclosures of each of which are herein incorporated by reference intheir entireties).

Antibody polypeptides of the invention having NGF binding specificitymay also be produced by constructing, using conventional techniques wellknown to those of ordinary skill in the art, an expression vectorcontaining an operon and a DNA sequence encoding an antibody heavy chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

A second expression vector is produced using the same conventional meanswell known to those of ordinary skill in the art, said expression vectorcontaining an operon and a DNA sequence encoding an antibody light chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

The expression vectors are transfected into a host cell by conventiontechniques well known to those of ordinary skill in the art to produce atransfected host cell, said transfected host cell cultured byconventional techniques well known to those of ordinary skill in the artto produce said antibody polypeptides.

The host cell may be co-transfected with the two expression vectorsdescribed above, the first expression vector containing DNA encoding anoperon and a light chain-derived polypeptide and the second vectorcontaining DNA encoding an operon and a heavy chain-derived polypeptide.The two vectors contain different selectable markers, but preferablyachieve substantially equal expression of the heavy and light chainpolypeptides. Alternatively, a single vector may be used, the vectorincluding DNA encoding both the heavy and light chain polypeptides. Thecoding sequences for the heavy and light chains may comprise cDNA,genomic DNA, or both.

The host cells used to express the antibody polypeptides may be either abacterial cell such as E. coli, or a eukaryotic cell. In a particularlypreferred embodiment of the invention, a mammalian cell of awell-defined type for this purpose, such as a myeloma cell, a Chinesehamster ovary (CHO) cell line, a NSO cell line, or a HEK293 cell linemay be used.

The general methods by which the vectors may be constructed,transfection methods required to produce the host cell and culturingmethods required to produce the antibody polypeptides from said hostcells all include conventional techniques. Although preferably the cellline used to produce the antibody is a mammalian cell line, any othersuitable cell line, such as a bacterial cell line such as an E.coli-derived bacterial strain, or a yeast cell line, may alternativelybe used.

Similarly, once produced the antibody polypeptides may be purifiedaccording to standard procedures in the art, such as for examplecross-flow filtration, ammonium sulphate precipitation, affinity columnchromatography and the like.

The antibody polypeptides described herein may also be used for thedesign and synthesis of either peptide or non-peptide mimetics thatwould be useful for the same therapeutic applications as the antibodypolypeptides of the invention. See, for example, Saragobi et al,Science, 253:792-795 (1991), the contents of which is hereinincorporated by reference in its entirety.

Screening Assays

The invention also includes screening assays designed to assist in theidentification of diseases and disorders associated with NGF in patientsexhibiting symptoms of an NGF associated disease or disorder.

In one embodiment of the invention, the anti-NGF antibodies of theinvention, or NGF binding fragments thereof, are used to detect thepresence of NGF in a biological sample obtained from a patientexhibiting symptoms of a disease or disorder associated with NGF. Thepresence of NGF, or elevated levels thereof when compared to pre-diseaselevels of NGF in a comparable biological sample, may be beneficial indiagnosing a disease or disorder associated with NGF.

Another embodiment of the invention provides a diagnostic or screeningassay to assist in diagnosis of diseases or disorders associated withNGF in patients exhibiting symptoms of an NGF associated disease ordisorder identified herein, comprising assaying the level of NGFexpression in a biological sample from said patient using apost-translationally modified anti-NGF antibody or binding fragmentthereof. The anti-NGF antibody or binding fragment thereof may bepost-translationally modified to include a detectable moiety such as setforth previously in the disclosure.

The NGF level in the biological sample is determined using a modifiedanti-NGF antibody or binding fragment thereof as set forth herein, andcomparing the level of NGF in the biological sample against a standardlevel of NGF (e.g., the level in normal biological samples). The skilledclinician would understand that some variability may exist betweennormal biological samples, and would take that into consideration whenevaluating results. In one embodiment of the invention, the anti-NGFantibodies of the invention may be used to correlate NGF expressionlevels with a particular stage of cancerous development. One skilled inthe art would be able to measure NGF in numerous subjects in order toestablish ranges of NGF expression that correspond to clinically definedstages of cancerous development. These ranges will allow the skilledpractitioner to measure NGF in a subject diagnosed with a cancer andcorrelate the levels in each subject with a range that corresponds to astage of said cancer. One skilled in the art would understand that bymeasuring NGF in the patient at different intervals, the progression ofthe cancer can be determined.

The above-recited assay may also be useful in monitoring a disease ordisorder, where the level of NGF obtained in a biological sample from apatient believed to have a NGF associated disease or disorder iscompared with the level of NGF in prior biological samples from the samepatient, in order to ascertain whether the NGF level in said patient haschanged with, for example, a treatment regimen.

The invention is also directed to a method of in vivo imaging whichdetects the presence of cells which express NGF comprising administeringa diagnostically effective amount of a diagnostic composition. Said invivo imaging is useful for the detection or imaging of NGF expressingtumors or metastases, for example, and can be useful as part of aplanning regimen for the design of an effective cancer treatmentprotocol. The treatment protocol may include, for example, one or moreof radiation, chemotherapy, cytokine therapy, gene therapy, and antibodytherapy, as well as an anti-NGF antibody or fragment thereof.

The present invention further provides for a kit for detecting bindingof an anti-NGF antibody of the invention to NGF. In particular, the kitmay be used to detect the presence of a NGF specifically reactive withan anti-NGF antibody of the invention or an immunoreactive fragmentthereof. The kit may also include an antibody bound to a substrate, asecondary antibody reactive with the antigen and a reagent for detectinga reaction of the secondary antibody with the antigen. Such a kit may bean ELISA kit and can comprise the substrate, primary and secondaryantibodies when appropriate, and any other necessary reagents such asdetectable moieties, enzyme substrates, and color reagents, for exampleas described herein. The diagnostic kit may also be in the form of animmunoblot kit.

A skilled clinician would understand that a biological sample includes,but is not limited to, sera, plasma, urine, saliva, mucous, pleuralfluid, synovial fluid and spinal fluid.

Methods of Ameliorating or Reducing Symptoms of, or Treating, orPreventing, Diseases and Disorders Associated with, NGF

In another embodiment of the invention, anti-NGF antibodies describedherein, or fragments thereof, i.e., preferably those which inhibit theassociation of NGF with TrkA and/or p75, are useful for ameliorating orreducing the symptoms of, or treating, or preventing, diseases anddisorders associated with NGF. Anti-NGF antibodies described herein, orfragments thereof, as well as combinations, can also be administered ina therapeutically effective amount to patients in need of treatment ofdiseases and disorders associated with NGF in the form of apharmaceutical composition as described in greater detail below.

In a preferred embodiment of the invention, the antibodies describedherein or fragments thereof, including Fab fragments, are utilized inmethods for the treatment of pain in a patient via administration ofsaid antibodies and/or fragments thereof.

In one embodiment of the invention, anti-NGF antibodies and/or fragmentsthereof described herein which inhibit the association of NGF with TrkAand/or p75, in conjunction with a second agent, are useful forameliorating or reducing the symptoms of, or treating, or preventing,the following non-limiting listing of diseases and disorders:inflammatory pain, post-operative incision pain, complex regional painsyndrome, cancer pain (particularly primary or metastatic bone cancerpain), fracture pain, osteoporotic fracture pain, pain resulting fromburn, osteoporosis, gout joint pain, pain associated with sickle cellcrises, and other nociceptic pain, as well as hepatocellular carcinoma,breast cancer, liver cirrhosis.

In another embodiment of the invention, anti-NGF antibodies and/orfragments thereof described herein, which inhibit the association of NGFwith TrkA and/or p75, in conjunction with a second agent, are useful forameliorating or reducing the symptoms of, or treating, or preventing,the following non-limiting listing of diseases and disorders:neurogenic, neuropathic or nociceptic pain. Neuropathic pain mayinclude, but is not limited to, trigeminal neuralgia, post-herpeticneuralgia, phantom limb pain, fibromyalgia, menstrual pain, ovarialgia,reflex sympathetic dystrophy and neurogenic pain. In other preferredembodiments, osteoarthritis or rheumatoid arthritis pain, lower backpain, diabetic neuropathy, sciatica, migraine, and other neuropathicpain.

Administration

In one embodiment of the invention, the anti-NGF antibodies describedherein, or NGF binding fragments thereof, which inhibit the associationof NGF with TrkA and/or p75, as well as combinations of said antibodiesor antibody fragments, and other anti-NGF antibodies or other activesare administered to a subject at a concentration of between about 0.1and 100.0 mg/kg of body weight of recipient subject. In a preferredembodiment of the invention, the anti-NGF antibodies described herein,or NGF binding fragments thereof, as well as combinations of saidantibodies or antibody fragments, are administered to a subject at aconcentration of about 0.4 mg/kg of body weight of recipient subject. Ina preferred embodiment of the invention, the anti-NGF antibodiesdescribed herein, or NGF binding fragments thereof, as well ascombinations of said antibodies or antibody fragments, are administeredto a recipient subject with a frequency of once every twenty-six weeksor less, such as once every sixteen weeks or less, once every eightweeks or less, once every four weeks or less, once every two weeks orless, once every week or less, or once daily or less.

Fab fragments may be administered every two weeks or less, every week orless, once daily or less, multiple times per day, and/or every fewhours. In one embodiment of the invention, a patient receives Fabfragments of 0.1 mg/kg to 40 mg/kg per day given in divided doses of 1to 6 times a day, or in a sustained release form, effective to obtaindesired results.

It is to be understood that the concentration of the antibody or Fabadministered to a given patient may be greater or lower than theexemplary administration concentrations set forth above.

A person of skill in the art would be able to determine an effectivedosage and frequency of administration through routine experimentation,for example guided by the disclosure herein and the teachings inGoodman, L. S., Gilman, A., Brunton, L. L., Lazo, J. S., & Parker, K. L.(2006). Goodman & Gilman's the pharmacological basis of therapeutics.New York: McGraw-Hill; Howland, R. D., Mycek, M. J., Harvey, R. A.,Champe, P. C., & Mycek, M. J. (2006). Pharmacology. Lippincott'sillustrated reviews. Philadelphia: Lippincott Williams & Wilkins; andGolan, D. E. (2008). Principles of pharmacology: the pathophysiologicbasis of drug therapy. Philadelphia, Pa., [etc.]: Lippincott Williams &Wilkins.

In another embodiment of the invention, the anti-NGF antibodiesdescribed herein, or NGF binding fragments thereof, as well ascombinations of said antibodies or antibody fragments, are administeredto a subject for treatment or prevention of pain and pain associatedconditions in a pharmaceutical formulation.

A “pharmaceutical composition” refers to a chemical or biologicalcomposition suitable for administration to a mammal. Such compositionsmay be specifically formulated for administration via one or more of anumber of routes, including but not limited to buccal, epicutaneous,epidural, inhalation, intraarterial, intracardial,intracerebroventricular, intradermal, intramuscular, intranasal,intraocular, intraperitoneal, intraspinal, intrathecal, intravenous,oral, parenteral, rectally via an enema or suppository, subcutaneous,subdermal, sublingual, transdermal, and transmucosal. In addition,administration can occur by means of injection, powder, liquid, gel,drops, or other means of administration.

In one embodiment of the invention, the anti-NGF antibodies describedherein, or NGF binding fragments thereof, preferably which inhibit theassociation of NGF with TrkA and/or p75, optionally in association withother antibodies and fragments thereof which inhibit the association ofNGF with TrkA as well as inhibiting the association of NGF with p75, andcombinations of said antibodies or antibody fragments, may be optionallyadministered in combination with one or more active agents includingother analgesic agents. Such active agents include analgesic,anti-histamine, antipyretic, anti-inflammatory, antibiotic, antiviral,and anti-cytokine agents. Active agents include agonists, antagonists,and modulators of TNF-α, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18,IFN-α, IFN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG, HepatocyteGrowth Factor (HGF), Hepcidin, including antibodies reactive against anyof the foregoing, and antibodies reactive against any of theirreceptors. Active agents also include but are not limited to2-Arylpropionic acids, Aceclofenac, Acemetacin, Acetylsalicylic acid(Aspirin), Alclofenac, Alminoprofen, Amoxiprin, Ampyrone, Arylalkanoicacids, Azapropazone, Benorylate/Benorilate, Benoxaprofen, Bromfenac,Carprofen, Celecoxib, Choline magnesium salicylate, Clofezone, COX-2inhibitors, Dexibuprofen, Dexketoprofen, Diclofenac, Diflunisal,Droxicam, Ethenzamide, Etodolac, Etoricoxib, Faislamine, fenamic acids,Fenbufen, Fenoprofen, Flufenamic acid, Flunoxaprofen, Flurbiprofen,Ibuprofen, Ibuproxam, Indometacin, Indoprofen, Kebuzone, Ketoprofen,Ketorolac, Lomoxicam, Loxoprofen, Lumiracoxib, Magnesium salicylate,Meclofenamic acid, Mefenamic acid, Meloxicam, Metamizole, Methylsalicylate, Mofebutazone, Nabumetone, Naproxen, N-Arylanthranilic acids,Nerve Growth Factor (NGF), Oxametacin, Oxaprozin, Oxicams,Oxyphenbutazone, Parecoxib, Phenazone, Phenylbutazone, Phenylbutazone,Piroxicam, Pirprofen, profens, Proglumetacin, Pyrazolidine derivatives,Rofecoxib, Salicyl salicylate, Salicylamide, Salicylates,Sulfinpyrazone, Sulindac, Suprofen, Tenoxicam, Tiaprofenic acid,Tolfenamic acid, Tolmetin, and Valdecoxib.

An anti-histamine can be any compound that opposes the action ofhistamine or its release from cells (e.g., mast cells). Anti-histaminesinclude but are not limited to acrivastine, astemizole, azatadine,azelastine, betatastine, brompheniramine, buclizine, cetirizine,cetirizine analogues, chlorpheniramine, clemastine, CS 560,cyproheptadine, desloratadine, dexchlorpheniramine, ebastine,epinastine, fexofenadine, HSR 609, hydroxyzine, levocabastine,loratidine, methscopolamine, mizolastine, norastemizole, phenindamine,promethazine, pyrilamine, terfenadine, and tranilast.

Antibiotics include but are not limited to Amikacin, Aminoglycosides,Amoxicillin, Ampicillin, Ansamycins, Arsphenamine, Azithromycin,Azlocillin, Aztreonam, Bacitracin, Carbacephem, Carbapenems,Carbenicillin, Cefaclor, Cefadroxil, Cefalexin, Cefalothin, Cefalotin,Cefamandole, Cefazolin, Cefdinir, Cefditoren, Cefepime, Cefixime,Cefoperazone, Cefotaxime, Cefoxitin, Cefpodoxime, Cefprozil,Ceftazidime, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftriaxone,Cefuroxime, Cephalosporins, Chloramphenicol, Cilastatin, Ciprofloxacin,Clarithromycin, Clindamycin, Cloxacillin, Colistin, Co-trimoxazole,Dalfopristin, Demeclocycline, Dicloxacillin, Dirithromycin, Doripenem,Doxycycline, Enoxacin, Ertapenem, Erythromycin, Ethambutol,Flucloxacillin, Fosfomycin, Furazolidone, Fusidic acid, Gatifloxacin,Geldanamycin, Gentamicin, Glycopeptides, Herbimycin, Imipenem,Isoniazid, Kanamycin, Levofloxacin, Lincomycin, Linezolid, Lomefloxacin,Loracarbef, Macrolides, Mafenide, Meropenem, Meticillin, Metronidazole,Mezlocillin, Minocycline, Monobactams, Moxifloxacin, Mupirocin,Nafcillin, Neomycin, Netilmicin, Nitrofurantoin, Norfloxacin, Ofloxacin,Oxacillin, Oxytetracycline, Paromomycin, Penicillin, Penicillins,Piperacillin, Platensimycin, Polymyxin B, Polypeptides, Prontosil,Pyrazinamide, Quinolones, Quinupristin, Rifampicin, Rifampin,Roxithromycin, Spectinomycin, Streptomycin, Sulfacetamide,Sulfamethizole, Sulfanilimide, Sulfasalazine, Sulfisoxazole,Sulfonamides, Teicoplanin, Telithromycin, Tetracycline, Tetracyclines,Ticarcillin, Tinidazole, Tobramycin, Trimethoprim,Trimethoprim-Sulfamethoxazole, Troleandomycin, Trovafloxacin, andVancomycin.

Active agents also include Aldosterone, Beclometasone, Betamethasone,Corticosteroids, Cortisol, Cortisone acetate, Deoxycorticosteroneacetate, Dexamethasone, Fludrocortisone acetate, Glucocorticoids,Hydrocortisone, Methylprednisolone, Prednisolone, Prednisone, Steroids,and Triamcinolone. Any suitable combination of these active agents isalso contemplated.

A “pharmaceutical excipient” or a “pharmaceutically acceptableexcipient” is a carrier, usually a liquid, in which an activetherapeutic agent is formulated. In one embodiment of the invention, theactive therapeutic agent is a humanized antibody for treatment orprevention of pain and pain associated conditions described herein, orone or more fragments thereof. The excipient generally does not provideany pharmacological activity to the formulation, though it may providechemical and/or biological stability, and release characteristics.Exemplary formulations can be found, for example, in Remington'sPharmaceutical Sciences, 19th Ed., Grennaro, A., Ed., 1995 which isincorporated by reference.

As used herein “pharmaceutically acceptable carrier” or “excipient”includes any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents that arephysiologically compatible. Specific examples include amino acids,carbohydrates, alcohols and salts commonly used for antibodyformulations, e.g., for intravenous or subcutaneous administration. Inone embodiment, the carrier is suitable for parenteral administration.Alternatively, the carrier can be suitable for intravenous,intraperitoneal, subcutaneous, intramuscular, or sublingualadministration. Pharmaceutically acceptable carriers include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. The use of such media and agents for pharmaceuticallyactive substances is well known in the art. Except insofar as anyconventional media or agent is incompatible with the active compound,use thereof in the pharmaceutical compositions of the invention iscontemplated. Supplementary active compounds can also be incorporatedinto the compositions.

Pharmaceutical compositions typically must be sterile and stable underthe conditions of manufacture and storage. The invention contemplatesthat the pharmaceutical composition is present in lyophilized form. Thecomposition can be formulated as a solution, microemulsion, liposome, orother ordered structure suitable to high drug concentration. The carriercan be a solvent or dispersion medium containing, for example, water,ethanol, polyol (for example, glycerol, propylene glycol, and liquidpolyethylene glycol), and suitable mixtures thereof. The inventionfurther contemplates the inclusion of a stabilizer in the pharmaceuticalcomposition. The proper fluidity can be maintained, for example, by theuse of a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.

In many cases, it will be preferable to include isotonic agents, forexample, sugars, polyalcohols such as mannitol, sorbitol, or sodiumchloride in the composition. Prolonged absorption of the injectablecompositions can be brought about by including in the composition anagent which delays absorption, for example, monostearate salts andgelatin. Moreover, the alkaline polypeptide can be formulated in a timerelease formulation, for example in a composition which includes a slowrelease polymer. The active compounds can be prepared with carriers thatwill protect the compound against rapid release, such as a controlledrelease formulation, including implants and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers(PLG). Many methods for the preparation of such formulations are knownto those skilled in the art.

For each of the recited embodiments, the compounds can be administeredby a variety of dosage forms. Any biologically-acceptable dosage formknown to persons of ordinary skill in the art, and combinations thereof,are contemplated. Examples of such dosage forms include, withoutlimitation, reconstitutable powders, elixirs, liquids, solutions,suspensions, emulsions, powders, granules, particles, microparticles,dispersible granules, cachets, inhalants, aerosol inhalants, patches,particle inhalants, implants, depot implants, injectables (includingsubcutaneous, intramuscular, intravenous, and intradermal), infusions,and combinations thereof.

The above description of various illustrated embodiments of theinvention is not intended to be exhaustive or to limit the invention tothe precise form disclosed. While specific embodiments of, and examplesfor, the invention are described herein for illustrative purposes,various equivalent modifications are possible within the scope of theinvention, as those skilled in the relevant art will recognize. Theteachings provided herein of the invention can be applied to otherpurposes, other than the examples described above.

These and other changes can be made to the invention in light of theabove detailed description. In general, in the following claims, theterms used should not be construed to limit the invention to thespecific embodiments disclosed in the specification and the claims.Accordingly, the invention is not limited by the disclosure, but insteadthe scope of the invention is to be determined entirely by the followingclaims.

The invention may be practiced in ways other than those particularlydescribed in the foregoing description and examples. Numerousmodifications and variations of the invention are possible in light ofthe above teachings and, therefore, are within the scope of the appendedclaims.

Certain teachings related to methods for obtaining a clonal populationof antigen-specific B cells were disclosed in U.S. Provisional patentapplication No. 60/801,412, filed May 19, 2006, the disclosure of whichis herein incorporated by reference in its entirety.

Certain teachings related to humanization of rabbit-derived monoclonalantibodies and preferred sequence modifications to maintain antigenbinding affinity were disclosed in International Application No.PCT/US2008/064421, corresponding to International Publication No.WO/2008/144757, entitled “Novel Rabbit Antibody Humanization Methods andHumanized Rabbit Antibodies”, filed May 21, 2008, the disclosure ofwhich is herein incorporated by reference in its entirety.

Certain teachings related to producing antibodies or fragments thereofusing mating competent yeast and corresponding methods were disclosed inU.S. patent application Ser. No. 11/429,053, filed May 8, 2006, (U.S.Patent Application Publication No. US2006/0270045), the disclosure ofwhich is herein incorporated by reference in its entirety.

Certain teachings related to anti-NGF compositions and uses thereof weredisclosed in U.S. provisional patent application No. 61/418,832, filedDec. 1, 2010, the disclosure of which is herein incorporated byreference in its entirety including the sequence listing.

Certain NGF antibody polynucleotides and polypeptides are disclosed inthe sequence listing accompanying this patent application filing, andthe disclosure of said sequence listing is herein incorporated byreference in its entirety.

The entire disclosure of each document cited (including patents, patentapplications, journal articles, abstracts, manuals, books, or otherdisclosures) in the Background of the Invention, Detailed Description,and Examples is herein incorporated by reference in their entireties.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the subject invention, and are not intended to limit thescope of what is regarded as the invention. Efforts have been made toensure accuracy with respect to the numbers used (e.g. amounts,temperature, concentrations, etc.) but some experimental errors anddeviations should be allowed for. Unless otherwise indicated, parts areparts by weight, molecular weight is average molecular weight,temperature is in degrees centigrade; and pressure is at or nearatmospheric.

EXAMPLES Example 1 Preparation of Antibodies that Bind NGF

By using the antibody selection protocol described herein, one cangenerate an extensive panel of antibodies.

Immunization Strategy

Rabbits were immunized with huNGF (R&D Systems, Minneapolis, Minn.).Immunization consisted of a first subcutaneous (sc) injection of 100 μgin complete Freund's adjuvant (CFA) (Sigma) followed by two boosts, twoweeks apart, of 50 μg each in incomplete Freund's adjuvant (IFA)(Sigma). Animals were bled on day 55, and serum titers were determinedby ELISA (antigen recognition) and by non-radioactive proliferationassay (Promega) using the T1165 cell line.

Antibody Selection Titer Assessment

To identify and characterize antibodies that bind to human NGF, antibodycontaining solutions were tested by ELISA. Briefly, neutravidin coatedplates (Thermo Scientific), were blocked with ELISA buffer (0.1 mg/mLBSA, 1×PBS pH 7.4, 0.002% Tween 20 and 0.005% sodium azide) for 1 hr atroom temperature. The plates were then coated with a 1 g/mL biotinylatedB-NGF solution in ELISA buffer for 1 hour at room temperature. This wasfollowed by a wash step (3× using PBS plus 0.05% Tween 20) and a secondblock with ELISA buffer. The recombinant antibodies were then added ontothe plates and incubated for 1 hour at room temperature and then washed3× with PBS/Tween solution. For development, an anti-rabbit Fc-HRP(1:5000 dilution in ELISA buffer) was added onto the wells and incubatedfor 45 min at RT. After a 3× wash step with PBS/Tween solution, theplate was developed using TMB substrate for 3 minutes, stopped using0.5M HCl and read at 450 nm.

Functional Titer Assessment

To test for the ability of NGF antibodies to block NGF-dependent cellproliferation, we used TF-1 cells (Chevalier et al. Expression andfunctionality of the TrkA proto-oncogene product/NGF receptor inundifferentiated hematopoietic cells. Blood (1994) vol. 83 (6) pp.1479-85). Briefly, TF-1 cells were maintained in 10% FBS cRPMI media(“complete media”) supplemented with rhuGM-CSF. On the day of the assay,the antibodies were serially diluted in complete media in a round bottom96 well plate. B-NGF (R&D systems) was concomitantly added and theresultant antibody/B-NGF mixture was incubated at 37° C. for 1 hr. Whilethe Ab and B-NGF mixture was incubating, TF-1 cells were washed 3× withcomplete media, counted and plated in a flat bottom 96 well plate using25,000 cells per well in a 50 L volume. After 1 hour incubation theNGF-Antibody mixtures were added onto the cells and the plates wereincubated for 48 hrs at 37° C. in a humidified 5% CO₂ incubator. Cellproliferation was measured using the “CellTiter” aqueous one solutioncell proliferation assay (Promega) according to the manufacturer'sinstructions. The dependency of the signals on the concentration ofantibody was analyzed, and IC50 values were calculated using theGraphPad Prism program.

Tissue Harvesting

Once acceptable titers were established, the rabbit(s) were sacrificed.Spleen, lymph nodes, and whole blood were harvested and processed asfollows:

Spleen and lymph nodes were processed into a single cell suspension bydisassociating the tissue and pushing through sterile wire mesh at 70 μm(Fisher) with a plunger of a 20 cc syringe. Cells were collected in PBS.Cells were washed twice by centrifugation. After the last wash, celldensity was determined by trypan blue. Cells were centrifuged at 1500rpm for 10 minutes; the supernatant was discarded. Cells wereresuspended in the appropriate volume of 10% dimethyl sulfoxide (DMSO,Sigma) in FBS (Hyclone) and dispensed at 1 ml/vial. Vials were stored at−70° C. in a slow freezing chamber for 24 hours and stored in liquidnitrogen.

Peripheral blood mononuclear cells (PBMCs) were isolated by mixing wholeblood with equal parts of the low glucose medium described above withoutFBS. 35 ml of the whole blood mixture was carefully layered onto 8 ml ofLympholyte Rabbit (Cedarlane) into a 45 ml conical tube (Corning) andcentrifuged 30 minutes at 2500 rpm at room temperature without brakes.After centrifugation, the PBMC layers were carefully removed using aglass Pasteur pipette (VWR), combined, and placed into a clean 50 mlvial. Cells were washed twice with the modified medium described aboveby centrifugation at 1500 rpm for 10 minutes at room temperature, andcell density was determined by trypan blue staining. After the lastwash, cells were resuspended in an appropriate volume of 10% DMSO/FBSmedium and frozen as described above.

B Cell Culture

On the day of setting up B cell culture, PBMC, splenocyte, or lymph nodevials were thawed for use. Vials were removed from LN2 tank and placedin a 37° C. water bath until thawed. Contents of vials were transferredinto 15 ml conical centrifuge tube (Corning) and 10 ml of modified RPMIdescribed above was slowly added to the tube. Cells were centrifuged for5 minutes at 1.5K rpm, and the supernatant was discarded. Cells wereresuspended in 10 ml of fresh media. Cell density and viability wasdetermined by trypan blue. Cells were washed again and resuspended at1E07 cells/80 μL, medium. Biotinylated huNGF (B huNGF) was added to thecell suspension at the final concentration of 3 ug/mL and incubated for30 minutes at 4° C. Unbound B huNGF was removed with two 10 ml washes ofphosphate-buffered (PBF):Ca/Mg free PBS (Hyclone), 2 mM ethylenediaminetetraacetic acid (EDTA), 0.5% bovine serum albumin (BSA) (Sigma-biotinfree). After the second wash, cells were resuspended at 1E07 cells/80 μlPBF. 20 μl of MACS® streptavidin beads (Milteni)/10E7 cells were addedto the cell suspension. Cells were incubated at 4° C. for 15 minutes.Cells were washed once with 2 ml of PBF/10E7 cells. After washing, thecells were resuspended at 1E08 cells/500 μl of PBF and set aside. AMACS® MS column (Milteni) was pre-rinsed with 500 ml of PBF on amagnetic stand (Milteni). Cell suspension was applied to the columnthrough a pre-filter, and unbound fraction was collected. The column waswashed with 1.5 ml of PBF buffer. The column was removed from the magnetstand and placed onto a clean, sterile 5 ml Polypropylene Falcon tube. 1ml of PBF buffer was added to the top of the column, and positiveselected cells were collected. The yield and viability of positive andnegative cell fraction was determined by trypan blue staining. Positiveselection yielded an average of 1% of the starting cell concentration.

A pilot cell screen was established to provide information on seedinglevels for the culture. Plates were seeded at 10, 25, 50, 100, or 200enriched B cells/well. In addition, each well contained 50K cells/wellof irradiated EL-4.B5 cells (5,000 Rads) and an appropriate level ofactivated rabbit T cell supernatant (See U.S. Patent ApplicationPublication No. 20070269868)(ranging from 1-5% depending on preparation)in high glucose modified RPMI medium at a final volume of 250 μl/well.Cultures were incubated for 5 to 7 days at 37° C. in 4% CO₂.

Identification of Selective Antibody Secreting B Cells

Cultures were tested for antigen recognition and functional activitybetween days 5 and 7.

Antigen Recognition Screening

The ELISA format used is as described above except 50 μl of supernatantfrom the B cell cultures (BCC) wells was used as the source of theantibody. The conditioned medium was transferred to antigen-coatedplates. After positive wells were identified, the supernatant wasremoved and transferred to a 96-well master plate(s). The originalculture plates were then frozen by removing all the supernatant except40 μl/well and adding 60 μl/well of 16% DMSO in FBS. Plates were wrappedin paper towels to slow freezing and frozen after the addition of 10%DMSO at −70° C.

Functional Activity Screening

To test for the ability of NGF antibodies to block NGF-dependent cellproliferation, we used TF-1 cells (Chevalier et al. Expression andfunctionality of the TrkA proto-oncogene product/NGF receptor inundifferentiated hematopoietic cells. Blood (1994) vol. 83 (6) pp.1479-85). Briefly, TF-1 cells were maintained in 10% FBS cRPMI media(“complete media”) supplemented with rhuGM-CSF. On the day of the assay,the antibodies were serially diluted in complete media in a round bottom96 well plate. B-NGF (R&D systems) was concomitantly added and theresultant antibody/B-NGF mixture was incubated at 37° C. for 1 hr. Whilethe Ab and B-NGF mixture was incubating, TF-1 cells were washed 3× withcomplete media, counted and plated in a flat bottom 96 well plate using25,000 cells per well in a 50 L volume. After 1 hour incubation theNGF-Antibody mixtures were added onto the cells and the plates wereincubated for 48 hrs at 37° C. in a humidified 5% CO₂ incubator. Cellproliferation was measured using the “CellTiter” aqueous one solutioncell proliferation assay (Promega) according to the manufacturer'sinstructions. The dependency of the signals on the concentration ofantibody was analyzed, and IC50 values were calculated using theGraphPad Prism program.

B Cell Recovery

Plates containing wells of interest were removed from −70° C., and thecells from each well were recovered with 5-200 μl washes of medium/well.The washes were pooled in a 1.5 ml sterile centrifuge tube, and cellswere pelleted for 2 minutes at 1500 rpm.

The tube was inverted, the spin repeated, and the supernatant carefullyremoved. Cells were resuspended in 100 μl/tube of medium. 100 μlbiotinylated NGF coated streptavidin M280 dynabeads (Invitrogen) and 16μl of goat anti-rabbit H&L IgG-FITC diluted 1:100 in medium was added tothe cell suspension.

20 μl of cell/beads/FITC suspension was removed, and 5 μl droplets wereprepared on a glass slide (Corning) previously treated with Sigmacote(Sigma), 35 to 40 droplets/slide. An impermeable barrier of paraffin oil(JT Baker) was added to submerge the droplets, and the slide wasincubated for 90 minutes at 37° C., 4% CO₂ in the dark.

Specific B cells that produce antibody can be identified by thefluorescent ring around them due to antibody secretion, recognition ofthe bead-associated biotinylated antigen, and subsequent detection bythe fluorescent-IgG detection reagent. Once a cell of interest wasidentified, the cell in the center of the fluorescent ring was recoveredvia a micromanipulator (Eppendorf). The single cell synthesizing andexporting the antibody was transferred into a 250 μl microcentrifugetube and placed in dry ice. After recovering all cells of interest,these were transferred to −70° C. for long-term storage.

Isolation of Antibody Sequences from Antigen-Specific B Cell

Antibody sequences were recovered using a combined RT-PCR based methodfrom a single isolated B-cell or an antigenic specific B cell isolatedfrom the clonal B cell population. Primers are designed to anneal inconserved and constant regions of the target immunoglobulin genes (heavyand light), such as rabbit immunoglobulin sequences, and a two-stepnested PCR recovery step is used to obtain the antibody sequence.Amplicons from each well are analyzed for recovery and size integrity.The resulting fragments are then digested with AluI to fingerprint thesequence clonality. Identical sequences display a common fragmentationpattern in their electrophoretic analysis. The original heavy and lightchain amplicon fragments are then restriction enzyme digested withHindIII and XhoI or HindIII and BsiwI to prepare the respective piecesof DNA for cloning. The resulting digestions are then ligated into anexpression vector and transformed into bacteria for plasmid propagationand production. Colonies are selected for sequence characterization.

Recombinant Production of Monoclonal Antibody of Desired AntigenSpecificity and/or Functional Properties

Correct full-length antibody sequences for each well containing a singlemonoclonal antibody are established and miniprep DNA is prepared usingQiagen solid-phase methodology. This DNA is then used to transfectmammalian cells to produce recombinant full-length antibody. Eitherantibody containing supernatants or protein-A affinity purifiedantibodies are tested for antigen recognition and functional propertiesto confirm the original characteristics are found in the recombinantantibody protein.

Antigen Specific ELISA

To identify and characterize antibodies and Fab fragments that bind tohuman NGF, antibody- and Fab-containing solutions were tested by ELISA.Briefly, neutravidin coated plates (Thermo Scientific), were blockedwith ELISA buffer (0.1 mg/mL BSA, 1×PBS pH 7.4, 0.002% Tween 20 and0.005% sodium azide) for 1 hr at room temperature. The plates were thencoated with a 1 g/mL biotinylated B-NGF solution in ELISA buffer for 1hour at room temperature. This was followed by a wash step (3× using PBSplus 0.05% Tween 20) and a second block with ELISA buffer. Therecombinant antibodies or Fabs were then added onto the plates andincubated for 1 hour at room temperature and then washed 3× withPBS/Tween solution. For development, an anti-human Fc-HRP or ananti-human Fab-fragment HRP (1:5000 dilution in ELISA buffer) was addedonto the wells and incubated for 45 min at RT. After a 3× wash step withPBS/Tween solution, the plate was developed using TMB substrate for 3minutes, stopped using 0.5M HCl, and read at 450 nm.

Results: FIGS. 24-40 demonstrate that anti-NGF antibodies Ab1-Ab21 bindto NGF. Furthermore, FIGS. 28 and 29 demonstrate that Fab antibodyfragments Fab1 and Fab2 bind to NGF.

Functional Activity Screening

To test for the ability of NGF antibodies to block NGF-dependent andTrkA receptor-mediated cell proliferation activity, we used TF-1 cells(Chevalier et al. Expression and functionality of the TrkAproto-oncogene product/NGF receptor in undifferentiated hematopoieticcells. Blood (1994) vol. 83 (6) pp. 1479-85). Briefly, TF-1 cells weremaintained in 10% FBS cRPMI media (“complete media”) supplemented withrhuGM-CSF. On the day of the assay, the antibodies were serially dilutedin complete media in a round bottom 96 well plate. B-NGF (R&D systems)was concomitantly added and the resultant antibody/B-NGF mixture wasincubated at 37° C. for 1 hr. While the Ab and B-NGF mixture wasincubating, TF-1 cells were washed 3× with complete media, counted andplated in a flat bottom 96 well plate using 25,000 cells per well in a50 L volume. After 1 hour incubation the NGF-Antibody mixtures wereadded onto the cells and the plates were incubated for 48 hrs at 37° C.in a humidified 5% CO₂ incubator. Cell proliferation was measured usingthe “CellTiter” aqueous one solution cell proliferation assay (Promega)according to the manufacturer's instructions. The dependency of thesignals on the concentration of antibody was analyzed, and IC50 valueswere calculated using the GraphPad Prism program.

Results: FIGS. 41-52 demonstrate that anti-NGF antibodies Ab1-Ab20inhibit the proliferation of TF-1 cells. Furthermore, FIG. 44demonstrates that Fab antibody fragments also inhibit the proliferationof TF-1 cells. These Fab antibody fragments were produced by: 1.) Pichiapastoris expression of Fab2; and 2.) enzymatic digestion of Ab21produced in Pichia pastoris (Fab1).

Example 2: Enzymatic Production of Fab Fragments

Papain digestions were conducted using immobilized papain(Thermo/Pierce) as per manufacturer's instructions. Briefly, purifiedantibodies were incubated in a cystein/HCl-containing buffer withimmobilized papain at 37° C. with gentle rocking. The digestion wasmonitored by taking an aliquot and analyzing using SDS-PAGE for cleavageof the heavy chain. To stop the reaction, the immobilized papain wasspun out and washed using 50 mM Tris pH 7.5 and filtered. Undigestedfull length antibody and Fc fragments were removed by using aMabSelectSure (GE) column.

Example 3 Yeast Cell Expression

Antibody genes: Genes were cloned and constructed that directed thesynthesis of a chimeric humanized rabbit monoclonal antibody.

Methods

Construction of Pichia pastoris Expression Vectors for Heavy and LightChain Antibodies.

The light and heavy chain fragments (chimera or humanized) werecommercially synthesized and subcloned into a pGAP expression vector.The pGAP expression vector uses the GAP promoter to drive expression ofthe immunoglobulin chain and the human serum albumin (HAS) leadersequence for export. In addition, this vector contains common elementssuch as a bacterial origin of replication, and a copy of the Sh ble genewhich confers resistance to the antibiotic Zeocin™ (phleomycin). Zeocin™provides a means of selection for strains that contain the desiredexpression vector integrated into their genome.

Transformation of Expression Vectors into Haploid met1 and lys3 HostStrains of Pichia pastoris

All methods used for transformation of haploid P. pastoris strains andmanipulation of the P. pastoris sexual cycle were done as described inPichia Protocols (Methods in Molecular Biology Higgings, D R, and Cregg,J M, Eds. 1998. Humana Press, Totowa, N.J.). Prior to transformationeach vector was linearized within the GAP promoter sequences to directthe integration of the vector into the GAP promoter locus of the P.pastoris genome. Haploid strains were transfected using electroporationand successful transformants were selected on YPD Zeocin™ plates andthen cultured in 96-well plates for two days. Haploid strains were matedand selected for their ability to grow in the absence of the auxotrophmarkers (i.e., Lys and Met). Diploid strains were then selected fortheir ability to express either full length or Fab antibody fragmentsusing a ForteBio Octet system fitted with Protein A biosensors tomonitor expression.

Example 4 Expression of Ab21 and Fab2 in Pichia pastoris

Two Pichia strains for expression of either full length Ab21 or Fab2antibody fragment were made. For both the full length or the Fabexpressing strains, haploids strains were created and subsequentlymated. One haploid strain expressed full length light sequences for Ab21and another haploid strain expressed either the full length Ab21 or atruncated form of heavy chain to express an Fab fragment (e.g., Fab2).Each diploid strain was used to generate a research cell bank and usedfor expression in a bioreactor.

First an inoculum was expanded using the research cell bank using mediumcomprised of the following nutrients (% w/v): yeast extract 3%,anhydrous dextrose 4%, YNB 1.34%, 0.004% Biotin with 100 mM potassiumphosphate. The culture was expanded for approximately 24 hours in ashaking incubator at 30° C. and 300 rpm to generate the inoculum for thefermenters. A 10% inoculum was then added to Labfors 2.5 L workingvolume vessels containing sterile growth medium. The growth medium forthe full length Ab21 was comprised of the following nutrients: potassiumsulfate 18.2 g/L, ammonium phosphate monobasic 36.4 g/L, potassiumphosphate dibasic 12.8 g/L, magnesium sulfate heptahydrate 3.72 g/L,sodium citrate dihydrate 10 g/L, glycerol 40 g/L, yeast extract 30 g/L,PTM1 trace metals 4.35 mL/L, and antifoam 204 1.67 mL/L. The PTM1 tracemetal solution was comprised of the following components: cupric sulfatepentahydrate 6 g/L, sodium iodide 0.08 g/L, manganese sulfate hydrate 3g/L, sodium molybdate dihyrate 0.2 g/L, boric acid 0.02 g/L, cobaltchloride 0.5 g/L, zinc chloride 20 g/L, ferrous sulfate heptahydrate 65g/L, biotin 0.2 g/L, and sulfuric acid 5 mL/L.

The growth medium for the Fab2 fragment was comprised of the followingnutrients: potassium sulfate 10.92 g/L, ammonium phosphate monobasic21.84 g/L, potassium phosphate dibasic 7.68 g/L, magnesium sulfateheptahydrate 3.72 g/L, sodium citrate dihydrate 10 g/L, glycerol 40 g/L,yeast extract 30 g/L, PTM1 trace metal solution 2.61 mL/L, and antifoam204 1.67 mL/L. The PTM1 trace metal solution was comprised of thefollowing components: cupric sulfate pentahydrate 6 g/L, sodium iodide0.08 g/L, manganese sulfate hydrate 3 g/L, sodium molybdate dihyrate 0.2g/L, boric acid 0.02 g/L, cobalt chloride 0.5 g/L, zinc chloride 20 g/L,ferrous sulfate heptahydrate 65 g/L, biotin 0.2 g/L, and sulfuric acid 5mL/L. Both proteins were expressed under similar conditions. Briefly,the bioreactor process control parameters were set as follows: Agitation1000 rpm, airflow 1.35 standard liter per minute, temperature 28° C. andpH was controlled at six using ammonium hydroxide. No oxygensupplementation was provided.

The fermentation cultures were grown for approximately 12 to 16 hoursuntil the initial glycerol was consumed as denoted by a dissolved oxygenspike. The cultures were starved for approximately three hours after thedissolved oxygen spike. After this starvation period, a bolus additionof ethanol was added to the reactor to reach 1% ethanol (w/v). Thefermentation cultures were allowed to equilibrate for 15 to 30 minutes.Feed addition was initiated 30 minutes post-ethanol bolus and set at aconstant rate of 1 mL/min for 40 minutes, then the feed pump wascontrolled by an ethanol sensor keeping the concentration of ethanol at1% for the remainder of the run. The feed was comprised of the followingcomponents: yeast extract 50 g/L, dextrose 500 g/L, magnesium sulfateheptahydrate 3 g/L, and PTM1 trace metals 12 mL/L. For fermentation ofthe full length Ab21, sodium citrate dihydrate (0.5 g/L) was also addedto the feed. The total fermentation time was approximately 90 hours.

Example 5 Inhibition of NGF-p75 Interactions

NGF is reported to interact with two receptors on the cell surface: TrkAand p75. A biolayer interferometry assay via the “Octet” was used tocharacterize the ability of anti-NGF antibodies to inhibit NGF-p75interactions. Briefly, streptavidin (SA) sensors were pre-wetted in 1×kinetics buffer (1×PBS ph7.4, 0.002% Tween 20, 0.005% sodium azide and0.1 mg/mL BSA). A baseline was obtained using again 1× kinetics buffer,followed by binding of the biotinylated antibody being tested andanother short baseline in 1× kinetics buffer. NGF (1 g/mL) was loadednext and the sensor was then transferred onto 1× kinetics buffer. Afterloading of NGF onto the antibody, on one sensor, all possible sites ofNGF were blocked using an un-labeled solution of the biotinylatedantibody at 5 g/mL. As control, a parallel sensor was submerged into 1×kinetics buffer during this second blocking step. Both sensors were thenexposed to a solution containing p75 (1.2 g/mL). The ability of anantibody to block NGF-p75 interactions was then characterized bymonitoring the increase in signal when antibody-immobilized NGF wasexposed to soluble p75.

Results: FIGS. 53 and 54 demonstrates that anti-NGF antibodies Ab3, Ab4,Ab15, and Ab16 do not inhibit binding of NGF to p75, while FIG. 55demonstrates that antibody Ab5 inhibits binding of NGF to p75.

Example 6 Neurite PC12 Assay

The ability of anti-NGF antibodies to block NGF signaling mediatedthrough the p75 and TrkA receptors was measured in vitro using a ratadrenal medulla cell line, PC12. PC12 cells express both p75 and TrkAreceptors on their cell surface (Urdiales et al. Cell cyclephase-specific surface expression of nerve growth factor receptors TrkAand p75(NTR). J Neurosci (1998) vol. 18 (17) pp. 6767-75); (Greene andTischler. Establishment of a noradrenergic clonal line of rat adrenalpheochromocytoma cells which respond to nerve growth factor. Proc NatlAcad Sci USA (1976) vol. 73 (7) pp. 2424-8). Briefly, PC12 cells weremaintained in culture using 15% FBS RPMI and grown on a collagenI-coated flask for 48 hours before priming. The cells were then ‘primed’for 72 hours by exposing them to 100 ng/mL NGF in differentiation media(1% horse serum RPMI). On the day of the assay, the cells were harvestedwith a cell scraper, resuspended, rinsed in differentiation media(without NGF) and plated onto a collagen I-coated 24-well plate. Thefinal concentration of NGF in the assay was 100 ng/mL. The antibodiesbeing tested were pre-incubated with the NGF at different molar ratios(from 10× to 0.1×) for 1 hour in differentiation media prior to addingthem onto the PC-12 cells. On day 3, the media was gently removed andantibody-NGF mixtures were replaced. On day 10, the wells were observedunder a microscope and representative fields were digitized using a 10×magnification lens.

Results: FIGS. 56-69 and FIGS. 78 and 79 demonstrate that anti-NGFantibodies Ab1-Ab3, Ab5-Ab11, Ab13, Ab15, Ab16, and Ab17-Ab19 inhibitthe outgrowth of PC-12 neurite cells at increasing concentrations. Itcan be seen that antibodies Ab3, Ab15 and Ab16, when assayed at the sameantibody concentrations as the other tested anti-NGF antibodies, showedsignificantly less inhibition of the outgrowth of PC-12 neurite cells.This difference is believed to be attributable to the fact that Ab3, Aband Ab16, all inhibit TrkA/NGF interactions and not NGF/p75interactions, whereas the remaining tested antibodies inhibit theinteraction of NGF with both TrkA and p75.

Example 7 Modulation of Pain Assessed by Gait Analysis

To assess the effect of anti-NGF agents (full length and Fab fragments)in their ability to modulate pain, a PGPS (peptidoglycanpolysaccharide)-induced arthritis model was used. Briefly, male Lewisrats were injected with a solution of PGPS into their right ankle on day(−)17. One day later, ankles were evaluated for an inflammatory responseto the PGPS injection and non-responders were eliminated. Responderswere allowed to recover for seventeen days before an IV tail veinreactivation with PGPS.

Full-length antibodies were dosed once, either 2 hours or the nightbefore reactivation. Fab fragments were administered once a day with thefirst dose administered two hours prior to reactivation. Gait analysiswas performed by applying ink to the ventral surface of the foot anddocumenting weight bearing during movement (footprints) across paper.The rear feet of the rats were placed in blue colored ink, and black inkwas applied to the dorsal side of the foot on the suspected painful leg.Rats were placed on paper and allowed to walk. Gaits were scored asfollows: 0=normal, equal ink staining on both feet; 1=slight limp, toestaining evident and some heel staining; 2=limping, toes only staining;3=dragging/carrying leg, black drag marks from dorsal side of footpresent; 4=carrying leg, no staining from painful leg.

Results: FIG. 70 demonstrates a statistically significant reduction inpain as assessed by Gait analysis following administration of antibodiesAb2, Ab6, and Ab8, when compared with results obtained with thecontrols.

FIG. 71 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab6 andFab1, when compared with results obtained with the controls.

FIG. 72 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab3, whencompared with results obtained with the controls.

FIG. 73 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab6 andantibody Ab21, when compared with results obtained with the controls.

Example 8 Inflammation in PGPS-Induced Arthritis

The PGPS (peptidoglycan polysaccharide) induced arthritis model used toassess pain (Example 7) also has an associated inflammation response. Toassess inflammation, all animals had caliper measurements taken of theirankles prior to reactivation on day 0, and then on days 1, 2, 3 and 4 todetermine any anti-inflammatory or pro-inflammatory effects present intreated rats.

Results: FIG. 74 demonstrates an increase in inflammation followingadministration of each of antibodies Ab2, Ab6, and Ab8, when comparedwith inflammation results for the controls.

FIG. 75 demonstrates no significant increase in inflammation followingadministration of the Fab1 antibody fragment, when compared withinflammation results for the control. In contrast, administration ofantibody Ab6 resulted in increased inflammation, when compared withinflammation results for the controls.

FIG. 76 demonstrates an increase in inflammation followingadministration of antibody Ab3, when compared with inflammation resultsfor the controls.

FIG. 77 also demonstrates an increase in inflammation followingadministration of antibody Ab6 and antibody Ab21, when compared withinflammation results for the controls.

Example 9

To assess the effect of anti-NGF agents (full length and Fab fragments)in their ability to modulate pain, a PGPS (peptidoglycanpolysaccharide)-induced arthritis model was used. Briefly, male Lewisrats were injected with a solution of PGPS into their right ankle on day(−)17. One day later, ankles were evaluated for an inflammatory responseto the PGPS injection and non-responders were eliminated. Responderswere allowed to recover for seventeen days before an IV tail veinreactivation with PGPS. On day 2 post-reactivation, animals were testedby gait for pain and randomized based on their pain response. Theanimals were then dosed via IV injection receiving 5 mg/kg of either anegative control antibody or a test agent.

Full-length antibodies and Fab fragments were dosed once, either 2 hoursor the night before reactivation. Gait analysis was performed byapplying ink to the ventral surface of the foot and documenting weightbearing during movement (footprints) across paper. The rear feet of therats were placed in blue colored ink, and black ink was applied to thedorsal side of the foot on the suspected painful leg. Rats were placedon paper and allowed to walk. Gaits were scored as follows: 0=normal,equal ink staining on both feet; 1=slight limp, toe staining evident andsome heel staining; 2=limping, toes only staining; 3=dragging/carryingleg, black drag marks from dorsal side of foot present; 4=carrying leg,no staining from painful leg.

Results: FIG. 80 demonstrates no significant change in overall wellness,as determined by body weight, following administration of antibody Ab3or Ab15, when compared with the change in body weight for the noreactivation control. In contrast, administration of negative controlantibody resulted in a reduction in body weight, when compared with thechange in body weight for the no reactivation control.

FIG. 81 demonstrates a statistically significant reduction in pain asassessed by Gait analysis following administration of antibody Ab3 orantibody Ab15, when compared with results obtained with the controlsfollowing example 9. In particular, a demonstrated statisticallysignificant reduction in pain at 72 hours post-reactivation as assessedby Gait analysis following administration of antibody Ab3 or antibodyAb15, when compared with results obtained with the controls followingexample 9.

1. A method of treating pain or eliciting an analgesic effect in anindividual, comprising administering an effective amount of at least oneanti-human NGF antibody or fragment thereof, wherein the anti-NGFantibody or antibody fragment selected from Ab1, Ab2, Ab3, Ab4, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17,Ab18, Ab19, Ab20, or Ab21, or an anti-human NGF antibody or antibodyfragment comprising the same CDRs as any of said anti-human NGFantibodies or antibody fragments. 2-118. (canceled)
 119. An anti-humanNGF antibody or fragment thereof, wherein the anti-NGF antibody orantibody fragment selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8,Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20,or Ab21 or an anti-NGF antibody or antibody fragment comprising the sameCDRs as any of the foregoing.
 120. A composition comprising (i) ananti-human NGF antibody or fragment thereof, wherein the anti-NGFantibody or antibody fragment is selected from Ab1, Ab2, Ab3, Ab4, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17,Ab18, Ab19, Ab20, or Ab21 or an anti-NGF antibody or antibody fragmentcomprising the same CDRs as any of the foregoing and (ii) apharmaceutically acceptable carrier.
 121. A vector or nucleic acid whichcomprises a nucleic acid which encodes an anti-human NGF antibody orfragment thereof according to claim 119 selected from Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16,Ab17, Ab18, Ab19, Ab20, or Ab21 or an anti-NGF antibody or antibodyfragment comprising the same CDRs as any of the foregoing.
 122. A hostcell which expresses an anti-human NGF antibody or fragment thereofaccording to claim 119 selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19,Ab20, or Ab21 or an anti-NGF antibody or antibody fragment comprisingthe same CDRs as any of the foregoing.
 123. A method of producing ananti-human NGF antibody or fragment thereof, comprising culturing a hostcell according to claim 121 and isolating an anti-human NGF antibody orfragment thereof from the host cell or a culture containing.